بسم الله الرحمن الرحيم. Classification of Elements l.C,H,O,N,S 2. Ca,P,mg,Na,K,Cl(Nutritionally elements) 3.Cr,Co,Fe,Cu,Zn,Mn,Mo,Se,I,…. They are Trace.

الله الله بسم بسمالرحيم الرحيم الرحمن الرحمن

Classification of ElementsClassification of Elements

l.C,H,O,N,Sl.C,H,O,N,S2. Ca,P,mg,Na,K,Cl(Nutritionally 2. Ca,P,mg,Na,K,Cl(Nutritionally

elements)elements)3.Cr,Co,Fe,Cu,Zn,Mn,Mo,Se,I,…. They 3.Cr,Co,Fe,Cu,Zn,Mn,Mo,Se,I,…. They

are Trace elementsare Trace elements4.As,Ni,Si,V (their functions for human 4.As,Ni,Si,V (their functions for human

life has not been known)life has not been known)5.Al,Pb,Hg,Cd(toxic elements)5.Al,Pb,Hg,Cd(toxic elements)

SodiomSodiom

DistributionDistribution

50% exteracellular50% exteracellular

10% intracellular10% intracellular

40% connected to bone40% connected to bone

FunctionFunction

CatalyseCatalyse

Regulate osmotic pressureRegulate osmotic pressure

Regulate blood pHRegulate blood pH

Exteracellular electerolyteExteracellular electerolyte

Exitation of nerveExitation of nerve

The function of membraneThe function of membrane

Increase Na Increase Na (hypernateriemia)(hypernateriemia)

DiarheaDiarhea

NosiaNosia

Decrease ADH – Diabetes insipidusDecrease ADH – Diabetes insipidus

HYPERALDOSTRONISMHYPERALDOSTRONISM

Coshing syndromCoshing syndrom

NotritionNotrition

DehydrationDehydration

Decrease Na Decrease Na (hyponateriemia)(hyponateriemia)

HYPORALDOSTRONISMHYPORALDOSTRONISM

ADRENAL DEFICIENCYADRENAL DEFICIENCY

NephriteNephrite

PotasioPotasiomm

DistributionDistribution

Interacellular electrolyteInteracellular electrolyte

Na/K Interacellular = 1/10Na/K Interacellular = 1/10

Na/K Exteracellular = 28/10Na/K Exteracellular = 28/10

FunctionFunction

CatalysiseCatalysise

Protein synthesisProtein synthesis

Glycogen synthesisGlycogen synthesis

Regulate osmotic pressureRegulate osmotic pressure

Regulate blood pHRegulate blood pH

Interacellular electerolyteInteracellular electerolyte

Exitation of nerveExitation of nerve

The function of membraneThe function of membrane

HyperkalemiaHyperkalemia

DehydrationDehydration

KetoasidoseKetoasidose

AcidoseAcidose

HypokalemiaHypokalemia

NutritionNutrition

AlkaloseAlkalose

AldosteronismAldosteronism

Coshing syndromCoshing syndrom

calciumcalcium

DistributionDistribution

1kg total body(25mol)1kg total body(25mol)

99% hard tissue Ca10(PO4)6(OH)299% hard tissue Ca10(PO4)6(OH)2

1gr (25mmole) exteracellular1gr (25mmole) exteracellular

300mg(7.5mmole) blood300mg(7.5mmole) blood

The form of CaThe form of Ca

1- 45% connected protein(80% 1- 45% connected protein(80% connected to Alb and 20% Glubolin)connected to Alb and 20% Glubolin)

2-10% Complex with ligand (citrate, 2-10% Complex with ligand (citrate, lactate, phosphate and bicarbonate)lactate, phosphate and bicarbonate)

3-45% Ionized3-45% Ionized

FunctionFunction

1-Second messenger1-Second messenger

2-Bone2-Bone

3-Coagulation3-Coagulation

PhosphorPhosphor

DistributionDistribution

620g total body(20mol)620g total body(20mol)Phosphor exteracellular= intracellularPhosphor exteracellular= intracellularPhosphor intracellular is Phosphor intracellular is

organic(phospholipid,phosphoprotein)organic(phospholipid,phosphoprotein)85%Phosphor extracellular is hydroxy 85%Phosphor extracellular is hydroxy

apatiteapatite1gr (25mmole) exteracellular1gr (25mmole) exteracellular300mg(7.5mmole) blood300mg(7.5mmole) blood

اعمالاعمال

1-1- غشاء ساختمانی غشاء جزء ساختمانی جزء

2-2- سازی همانند و انرژی متابولیسم سازی در همانند و انرژی متابولیسم در

اکسیداتیو -3 -3 اکسیداتیو فسفریالسیون فسفریالسیون

4-4- شکستنگلیکوژن مانند واکنشها شکستنگلیکوژن در مانند واکنشها در

ساخت -5 -5 ساخت در دیفسفوگلیسرات دیفسفوگلیسرات 33--22در

6-6- آپاتیت هیدروکسی ساختمانی آپاتیت جزء هیدروکسی ساختمانی جزء

متابولیسم تنظیم در موثر متابولیسم هورمونهای تنظیم در موثر هورمونهایوفسفر وفسفر کلیسم کلیسم

پاراتیروئیدهورمونپاراتیروئیدهورمون-1-1

کلسیتریولکلسیتریول-2-2

کلسیتونینکلسیتونین-3-3

پاراتیروئیدهورمون پاراتیروئیدهورمون اعمال اعمال

وحرکت -1-1 استئوسیتها واحتماال استئوکالستها وحرکت تحریک استئوسیتها واحتماال استئوکالستها تحریک) ( کلسیتریول با سینرجیستیک اثر ازاستخوان (کلسیم ( کلسیتریول با سینرجیستیک اثر ازاستخوان کلسیم

2-2- کلسیم کلیوی بازجذب کلسیم افزایش کلیوی بازجذب افزایش

3-3- فسفات کلیوی بازجذب فسفات مهار کلیوی بازجذب مهار

واسیدهای -4 -4 سدیم،پتاسیم بیکربنات، کلیوی بازجذب واسیدهای مهار سدیم،پتاسیم بیکربنات، کلیوی بازجذب مهارآمینهآمینه

5-5- تبدیل ) تبدیل ) تحریک به 2525تحریک کلسیتریول به هیدروکسی کلسیتریول دی دی 2525وو11هیدروکسی) کلسیتریول (هیدروکسی کلسیتریول هیدروکسی

6-6- ای روده کلسیم جذب ای افزایش روده کلسیم جذب افزایش

کلسیتریولکلسیتریول

DD ویتامینویتامین است است دونوع ::دونوع

1- 1- D3D3 کلسیفرول کلسیفرول کله کله

2- 2- D2D2ارگوکلسیفرولارگوکلسیفرول

DD ویتامینویتامین 2525به به OHDOHD وبعد وبعد1,251,25بهبه (OH)2D(OH)2D((کلسیتریولکلسیتریول))

کلسیتریول کلسیتریول اعمال اعمال

1-1- روده از کلسیم جذب روده تحریک از کلسیم جذب تحریک

استخوان -2 -2 از کلسیم جذب استخوان تحریک از کلسیم جذب تحریک

3-3- کلیه از کلسیم کلیه بازجذب از کلسیم بازجذب

4-4- فسفر ای روده جذب فسفر افزلیش ای روده جذب افزلیش

کلسیتونینکلسیتونین

تیروئید سی تیروئید درسلولهای سی درسلولهای

جذب: وکاهش حلقوی پی ام افزایش جذب: عمل وکاهش حلقوی پی ام افزایش عملوفسفات وفسفات کلسیم کلسیم

کلسیم کلسیم افزایش افزایش

هیپرپاراتیروئیدیسمهیپرپاراتیروئیدیسم-1-1

2-2- دو هر یا کلسیم یا د ویتامین زیاد دو مصرف هر یا کلسیم یا د ویتامین زیاد مصرف

استخوانی، ) -3-3 کارسنومای استخوانی استخوانی، ) بیماریهای کارسنومای استخوانی بیماریهایاستئوپورز ، استئوپورز لوسمی ، لوسمی

4-4- هورمونها اکتوپیک هورمونها ترشح اکتوپیک ترشح

مانند -5-5 هیپرکلسیمی کننده القاء داروهای مانند مصرف هیپرکلسیمی کننده القاء داروهای مصرفدار کلسیم اسیدهای آنتی و دار تیازیدها کلسیم اسیدهای آنتی و تیازیدها

کلسیم کلسیم کاهش کاهش

هیپوپاراتیروئیدیسمهیپوپاراتیروئیدیسم-1-1

2-2-) استئوماالسی ) و راشیتیسم د (کمبودویتامین استئوماالسی ) و راشیتیسم د کمبودویتامین

3-3- کلیوی کلیوی نارسایی نارسایی

4-4- جذب سوء جذب سندرم سوء سندرم

5-5- خون پروتئین خون کاهش پروتئین کاهش

Trace elementTrace element

Definitions and ClssificationDefinitions and Clssification

• Are those elements with the blood Are those elements with the blood level of ug/L and tissue level of ug/g level of ug/L and tissue level of ug/g tissuetissue

• Daily consumption of are within Daily consumption of are within mg/daymg/day

CHARACTERISTICS OF CHARACTERISTICS OF TRACE ELEMENTSTRACE ELEMENTS l.Amplification:Small amounts need for l.Amplification:Small amounts need for

biochemical biochemical actions(HORMONES,ENZYMES,PROTEINS)actions(HORMONES,ENZYMES,PROTEINS)

2.Specificity.Essential trace elements are 2.Specificity.Essential trace elements are specific for their funtionsspecific for their funtions

They interact with electron They interact with electron donor(nitrogen,sulfur,oxygen)donor(nitrogen,sulfur,oxygen)

3.Homostasis.(absorption,storage,excretion)3.Homostasis.(absorption,storage,excretion) 4.Interaction.T.E interfers with each other from 4.Interaction.T.E interfers with each other from

their sites of absorption to the site of utilizationstheir sites of absorption to the site of utilizations Zn and Ca, Mo and Cu,….Zn and Ca, Mo and Cu,….

LABORATORY LABORATORY ASSESSMENTSASSESSMENTS

• Measurement of Measurement of metalloenzymemetalloenzyme

• Measurement of Measurement of metals in metals in biological fluidsbiological fluids

SAMPLE COLLECTIONSSAMPLE COLLECTIONS

• BLOODBLOOD

• SalivaSaliva

• SweetSweet

• HairHair

• NailNail

• UrineUrine

• etcetc

ANALYTICAL TECHNIQUESANALYTICAL TECHNIQUES

• Atomic aaaaaabsorption Atomic aaaaaabsorption Spectrophotometry(A.A.S,l955)Spectrophotometry(A.A.S,l955)

• Neutron Activation AnalysisNeutron Activation Analysis

• ColorimetryColorimetry

• Inductivity coupled plasmaInductivity coupled plasma

• Chemical and physiochemical methodsChemical and physiochemical methods

IronIron

Iron Metabolism in HumanIron Metabolism in Human

• Iron AbsorptionIron Absorption

• Blood circulationBlood circulation

• Intracellular movementIntracellular movement

• Heme synthesisHeme synthesis

آهن آهن منابع منابع

1-1- : تخم زرده گوشت، کلیه، کبد، منابع ترین : غنی تخم زرده گوشت، کلیه، کبد، منابع ترین غنیمرغمرغ

2-2- تازه: ماهی،سبزیجات عضالت، تازه: متوسط ماهی،سبزیجات عضالت، متوسط

3-3- : جات میوه اکثر و زمینی سیب برنج فقیر : منابع جات میوه اکثر و زمینی سیب برنج فقیر منابع

Iron AbsorptionIron Absorption

• Uptake of iron from lumenUptake of iron from lumen

• Transfer of iron across the cell Transfer of iron across the cell

• Exocytosis of ironExocytosis of iron

Factors Affectting Iron Factors Affectting Iron absorptionabsorption

• Amino acidsAmino acids

• Chelating agentsChelating agents

• Form of ironForm of iron

Principles of Iron Principles of Iron absorptionabsorption• Iron in diet(heme and none heme)Iron in diet(heme and none heme)• None heme iron binds strongly to proteins None heme iron binds strongly to proteins

and polysaccharidesand polysaccharides• Less than l0 percent of iron will be absorbedLess than l0 percent of iron will be absorbed• Iron absorption by intestine is an energy Iron absorption by intestine is an energy

dependent processdependent process• Vitamine C and stomach fluid fascilate iron Vitamine C and stomach fluid fascilate iron

absorptionabsorption• Erythropoitin seems to be important in iron Erythropoitin seems to be important in iron

absorptionabsorption

Increase iron absorptionIncrease iron absorption

• Iron deficiencyIron deficiency

• Pyridoxin deficiencyPyridoxin deficiency

• Late pregnancyLate pregnancy

• HemochromatosisHemochromatosis

• HypoxiaHypoxia

• Increase erythropoiesisIncrease erythropoiesis

Reduced iron absorptionReduced iron absorption

• Decrease erythropoiesisDecrease erythropoiesis

• Iron overloadIron overload

• Malignant inflammatoryMalignant inflammatory

• InfectionsInfections

• High phosphate and phytateHigh phosphate and phytate

• MalabsorptionMalabsorption

Distribution of ironDistribution of iron

• Red blood cell Hb 70.5percentRed blood cell Hb 70.5percent

• Muscle myoglobin 3.2 percentMuscle myoglobin 3.2 percent

• Store Ferritin 26 percentStore Ferritin 26 percent

• Blood Tf 0.l percentBlood Tf 0.l percent

• Various enzyme o.l percentVarious enzyme o.l percent

• CytochromesCytochromes

• CatalaseCatalase

•

Iron in BloodIron in Blood• Iron binds to transferrinIron binds to transferrin

•Transferrin is a beta glycoproteinTransferrin is a beta glycoprotein

• It has 679 amino acidsIt has 679 amino acids

•There are two iron binding sites on itThere are two iron binding sites on it

• It was discovered by Caroline in l944It was discovered by Caroline in l944

•Fontes and thivalle in 1925Fontes and thivalle in 1925

Human TransferrinHuman Transferrin

• SerotransferrinSerotransferrin

• SiderophyllinSiderophyllin

• Glycoprotein with 8-10 percent Glycoprotein with 8-10 percent carbohydratecarbohydrate

• Found in blood as Apotf,mono-ferric tfFound in blood as Apotf,mono-ferric tf

• And differric tfAnd differric tf

• Separated by Urea gel electrophoresisSeparated by Urea gel electrophoresis

Human TransferrinHuman Transferrin

• Synthetized by HepatocytesSynthetized by Hepatocytes• It is a single chain glycoproteinIt is a single chain glycoprotein• Molecular weight of 80kDMolecular weight of 80kD• PI=5.4-5.9PI=5.4-5.9• N-acetyllacosamine binds ti N-acetyllacosamine binds ti

Aspargin of TfAspargin of Tf• ASN 611 and ASN 413 are ligands ASN 611 and ASN 413 are ligands

close to C-terminalclose to C-terminal

The requirements for Iron The requirements for Iron binding to Tfbinding to Tf

• Fe + Citrate --Fe + Citrate --Fe-citrateFe-citrate

• Fe-citrate +Bicarbonate-Fe-citrate +Bicarbonate-Fe-Fe-bicarbonatebicarbonate

• Fe-bi +apotf-Fe-bi +apotf-Fe-bi apotfFe-bi apotf

• Fe +Citrate +bi + Apotf-Fe +Citrate +bi + Apotf-Tf Tf +citrate+citrate

• Fe>Cr>Cu> Mn> Co>NiFe>Cr>Cu> Mn> Co>Ni

Transferrin FunctionTransferrin Function

• Transfer of iron from site of Transfer of iron from site of absorption to the site of utilization absorption to the site of utilization in the cellin the cell

• Erythropoietic cellsErythropoietic cells

• Non-erythropoietic cellsNon-erythropoietic cells

• Transfer to hepatocytesTransfer to hepatocytes

• Involve in immuno systemInvolve in immuno system

Relationship between Relationship between transferrin and TIBCtransferrin and TIBC

• The number of binding sites on the The number of binding sites on the transferrintransferrin

• Transferrin(mg/dl)=0.7 x TIBCTransferrin(mg/dl)=0.7 x TIBC

• Transferrin saturation=l00 xserum Transferrin saturation=l00 xserum Fe/serum TIBCFe/serum TIBC

Intrcellular movement of Intrcellular movement of IronIron

Binding of Fe-transferrin to Binding of Fe-transferrin to the receptorsthe receptors

Endocytosis of of transferrin Endocytosis of of transferrin with ironwith iron

Release of iron from Release of iron from transferrintransferrin

Exocytosis of apo-tfExocytosis of apo-tf

Direct donation of iron from Direct donation of iron from transferrin to the transferrin to the mitochondriamitochondria

StorageStorage

1-Ferritine1-Ferritine

2-Hemosidrine(liver, spleen and bone 2-Hemosidrine(liver, spleen and bone marrow) unsolublemarrow) unsoluble

Iron DeficiencyIron Deficiency

• Iron deficiency may be due toIron deficiency may be due to

• Low diet ironLow diet iron

• Low iron absorptionLow iron absorption

• High excretion of iron High excretion of iron

• Therefor,Low plasma ironTherefor,Low plasma iron

• High plasma TIBCHigh plasma TIBC

• Low plasma FerritinLow plasma Ferritin

• Anemia is documentedAnemia is documented

Iron OverloadIron Overload

• Iron overload can be due to;Iron overload can be due to;

• High iron consumption(chronic or High iron consumption(chronic or acute)acute)

• TransfusionTransfusion

• HemochromatosisHemochromatosis

• ToxificationToxification

• HemosidrosisHemosidrosis

HemochromatosisHemochromatosis

• It is a rare diseaseIt is a rare disease• High iron absorptionHigh iron absorption• No iron absorption controlNo iron absorption control• Production of hemosiderin in skinProduction of hemosiderin in skin• Deposition of iron in liver causes cyrosisDeposition of iron in liver causes cyrosis• Damage to pancreatic function causes of Damage to pancreatic function causes of

diabetediabete• Myocardia infarctionMyocardia infarction• It is familial disorderIt is familial disorder

آهن وضعیت آهن بررسی وضعیت بررسی

سرم- : 11 آهن آهن افزایش سرم- : در آهن آهن افزایش دردسیلیتر،/ 200200<< دسیلیتر،/ میکروگرم ترانسفرین ترانسفرین TIBCTIBCمیکروگرم

آزاد%( 5555اشباع)<اشباع)< پورفیرین و فریتین آزاد%( البته پورفیرین و فریتین البتهنیست مناسب اریتروسیتها از نیست حاصل مناسب اریتروسیتها از حاصل

22 : مخصوصاپورفیرین- و فریتین آهن کاهش : در مخصوصاپورفیرین- و فریتین آهن کاهش دراست مناسب اریتروسیتها از حاصل است آزاد مناسب اریتروسیتها از حاصل آزاد

سرم آهن سرم افزایش آهن افزایش

هموسیدروزهموسیدروز -1 -1

هموکروماتوزهموکروماتوز-2-2

3-3- ( خونی کم تاالسمی، اریتروسیتها تشکیل ) کاهش خونی کم تاالسمی، اریتروسیتها تشکیل کاهش) پیریدوکسین کمبود اثر در (پرنیشیوز پیریدوکسین کمبود اثر در پرنیشیوز

4-4- همولیتیک بیماریهای در اریتروسیتها تجزیه همولیتیک افزایش بیماریهای در اریتروسیتها تجزیه افزایش

همولیزهمولیز-5-5

مصرفمصرف-6-6

سرم آهن سرم کاهش آهن کاهش

آهن -1 -1 فقر خونی آهن کم فقر خونی کم

وبیماریهای -2 -2 عفونت اثر در نرموکرمیک وبیماریهای آنمی عفونت اثر در نرموکرمیک آنمیمزمنمزمن

نفروزنفروز-3-3

مصرفمصرف-4-4

TIBCTIBC

آهن- 11افزایش:افزایش: فقر آهن- آنمی فقر ،استروژن- 22آنمی ،استروژن- حاملگی حاملگیخون- 33تراپی تراپی ومزمن حاد دادن خون- ازدست ومزمن حاد دادن --44ازدستحادکبدی حادکبدی آسیب آسیب

کبدی- 22هموکروماتوز-هموکروماتوز-11کاهش:کاهش: کبدی- سیروز تاالسمی- تاالسمی- 33سیروزمزمن- 44 بیماریهای و عفونت اثر در مزمن- آنمی بیماریهای و عفونت اثر در آنمی

) نئوپالسمها) از ( بعضی نئوپالسمها) از نفروز-نفروز-55بعضی

سرم سرم ترانسفرین ترانسفرین

آهن- 11افزایش:افزایش: فقر آهن- آنمی فقر ،استروژن- 22آنمی ،استروژن- حاملگی حاملگیتراپیتراپی

دراثر- 11کاهش:کاهش: میکروسیتیک هموکرمیک دراثر- آنمی میکروسیتیک هموکرمیک آنمیمزمن مزمن بیماریهای ازدست- 22بیماریهای یا پروتئین ازدست- نقصان یا پروتئین نقصان

نفروز ، مزمن سوختن،عفونت اثر در نفروز دادن ، مزمن سوختن،عفونت اثر در - - 33دادنژنتیکی ژنتیکی نقصان نقصان

CoppeCopperr

Contents Contents

• Introduction Introduction

• Copper uptake Copper uptake

• Copper distribution Copper distribution

• Copper related diseaseCopper related disease

Introduction Introduction

• Cu is the 29th element of the Periodic Table with Cu is the 29th element of the Periodic Table with electronic configuration 3d10 4s1electronic configuration 3d10 4s1

• Occurs in human and animal tissues in biological Occurs in human and animal tissues in biological systems in both the +1 and +2 valence statessystems in both the +1 and +2 valence states

• There are 80 mg copper in total in adult humans There are 80 mg copper in total in adult humans (higher concentration in liver and brain)(higher concentration in liver and brain)

• The major functions of copper–biological The major functions of copper–biological compounds involve oxidation–reduction compounds involve oxidation–reduction reactionsreactions

Why is copper essential for Why is copper essential for humans?humans?

• Copper deficiencyCopper deficiency– Connective tissue disordersConnective tissue disorders

•Cardiovascular conditions and Cardiovascular conditions and osteoporosisosteoporosis

– In infants causes iron-unresponsive In infants causes iron-unresponsive anemia, Neutropenia and bone anemia, Neutropenia and bone abnormalitiesabnormalities

Some copper enzymes in Some copper enzymes in humanszhumansz

All organisms require copper as a catalytic cofactor for biological processes such as

Respiration Iron transportOxidative stress protectionPeptide hormone productionPigmentation Blood clottingNormal cell growth and development

However,Copper also causes damage to

Lipids, proteins and DNACopper imbalances in humans lead to serious diseases

Menkes syndrome or Wilson disease

Inability to appropriately distribute copper to all cells and tissues

Neurodegenerative diseasesFamilial amyotropic lateral sclerosisAlzheimer’s disease Prion diseases

Cells have developed Dedicated components and sophisticated homeostatic mechanisms to acquire and maintain adequate intracellular copper concentrations

Absorption Copper is absorbed from the diet in the stomach(small intestine)Most likely as complex with amino acids (histidine or peptides)

In the plasma ≥ 90% of the total , Caeruloplasmin

Caeruloplasmin → copper-requiring cells (Caeruloplasmin receptors)

Only 5% is bound to albumin or to amino acids (histidine)

In the liverCopper is bound to protein caeruloplasmin

With six or seven copper ions per molecule

Pathway of copper in the Pathway of copper in the bodybody

• Copper distributionCopper distribution– Three independent copper-delivery Three independent copper-delivery

pathways:pathways:•Delivery to the secretory pathwayDelivery to the secretory pathway•To Cu,Zn SODTo Cu,Zn SOD•To COX in mitochondria To COX in mitochondria

•Delivery of copper to the secretory Delivery of copper to the secretory pathwaypathway– Clotting factors V and VIIIClotting factors V and VIII– Tyrosinase Tyrosinase – Lysyl oxidaseLysyl oxidase– Ceruloplasmin Ceruloplasmin – Fet3 multicopper ferroxidase Fet3 multicopper ferroxidase

•For high-affinity iron uptake in yeast For high-affinity iron uptake in yeast •Delivery of copper to the secretory Delivery of copper to the secretory

pathway:pathway:– Atx1 (or Atox1 in mammals)Atx1 (or Atox1 in mammals)– Copper-transporting ATPase Ccc2 (or Copper-transporting ATPase Ccc2 (or

ATP7A, ATP7B in mammals)ATP7A, ATP7B in mammals)

• Atx1/Atox1 and Ccc2/ATP7A/B proteins Atx1/Atox1 and Ccc2/ATP7A/B proteins directly interact, in a copper-stimulated directly interact, in a copper-stimulated manner manner – Metal is transferred via a series of ligand-Metal is transferred via a series of ligand-

exchange reactions involving two-and exchange reactions involving two-and three-coordinate intermediates between three-coordinate intermediates between cysteine ligands in the CxxC motifs cysteine ligands in the CxxC motifs

• The Ccc2/ATP7A/B copper pumpsThe Ccc2/ATP7A/B copper pumps– Contain between two and six of the Contain between two and six of the

MxCxxC metal binding domains (MBDs) at MxCxxC metal binding domains (MBDs) at their amino termini their amino termini

– Each MBD binds a single copper atomEach MBD binds a single copper atom

• A positively charged surface on Atx1/Atox1 is A positively charged surface on Atx1/Atox1 is thought to engage in ionic interactions with thought to engage in ionic interactions with negatively charged residues on the MBDs of copper negatively charged residues on the MBDs of copper pumps pumps

• Under low copper conditionsUnder low copper conditions– Both proteins localized in the trans-Golgi network Both proteins localized in the trans-Golgi network

•Deliver copper to secreted cupro-proteins Deliver copper to secreted cupro-proteins

• Elevated copper levelsElevated copper levels– ATP7A,re-distributes to plasma membraneATP7A,re-distributes to plasma membrane

• ATP7A ATP7A – Contains a di-leucine motif near the Contains a di-leucine motif near the

carboxyl terminus carboxyl terminus – Mediates its endocytosis back to the Mediates its endocytosis back to the

Golgi Golgi • ATP7BATP7B

– Expressed largely in the liver and in Expressed largely in the liver and in neuronal tissueneuronal tissue•Undergoes copper-induced trafficking Undergoes copper-induced trafficking from the trans-Golgi network to from the trans-Golgi network to intracellular vesicles intracellular vesicles

•Copper excretion from the liver to the Copper excretion from the liver to the bilebile

• ATP7A and ATP7B have six MBDs within ATP7A and ATP7B have six MBDs within the amino-terminal regionthe amino-terminal region

Two-dimensional Two-dimensional representation of the Menkes representation of the Menkes and Wilson Cu-ATPases and Wilson Cu-ATPases

Copper-induced trafficking of the Copper-induced trafficking of the Menkes (a) and Wilson (b) Cu-Menkes (a) and Wilson (b) Cu-ATPaseATPase

The cellular pathways of The cellular pathways of coppercopper

Copper as a catalystCopper as a catalyst

• Cu(II) catalyses efficient formation Cu(II) catalyses efficient formation of 8-aminodeoxyguanosine as well of 8-aminodeoxyguanosine as well as 8-oxodGas 8-oxodG

• Copper(II) forms thioxyl radicals, Copper(II) forms thioxyl radicals, RS˚, which may produce RSSR,that RS˚, which may produce RSSR,that can be damaging → Conformational can be damaging → Conformational changes (misfolding) changes (misfolding)

Concentrations above the trace essential metal

Copper ions exposed to oxidants

H2O2 inactivate CuZn-SOD(oxidation of the histidine active site)

loss of copper ion

Damage to DNAstrand breakshydroxylated base products Cancer

ROS

• Copper(II) compounds induce vomiting in Copper(II) compounds induce vomiting in human beings human beings – An emetic for general poisoningAn emetic for general poisoning

• Copper was found to bind DNA with high Copper was found to bind DNA with high affinity. copper binds to N7 of guanine affinity. copper binds to N7 of guanine residue and forming a pseudo-octahedral residue and forming a pseudo-octahedral geometry in which the other sites are geometry in which the other sites are occupied with water moleculesoccupied with water molecules

• The binding of copper ions to specific sites The binding of copper ions to specific sites can modify the conformational structures of can modify the conformational structures of proteins, polynucleotides or DNA and proteins, polynucleotides or DNA and biomembranesbiomembranes

Diseases resulting from Diseases resulting from mutations in genes encoding mutations in genes encoding copper transport proteinscopper transport proteins

Wilson’s diseaseWilson’s disease

• Metal storage disorderMetal storage disorder• An autosomal recessive An autosomal recessive

– incidenceincidence•one in 35 000 to 100 000 one in 35 000 to 100 000

– Biliary excretion of the metal ion is defectiveBiliary excretion of the metal ion is defective•Build up of copper, death of hepatocytes, release Build up of copper, death of hepatocytes, release

of copper, accumulation in extra-hepatic tissuesof copper, accumulation in extra-hepatic tissues• Low level of plasma holoceruloplasmin Low level of plasma holoceruloplasmin • Ceruloplasmin protein concentrations may be close Ceruloplasmin protein concentrations may be close

to normal to normal

Wilson’s diseaseWilson’s disease

•The primary genetic defect in The primary genetic defect in Wilsons disease is in ATP7B Wilsons disease is in ATP7B – predominately located at the predominately located at the

trans-Golgi networktrans-Golgi network

•Major contribution to the pathology Major contribution to the pathology is Cu-mediated oxidative damageis Cu-mediated oxidative damage– activation of cell death pathwaysactivation of cell death pathways

Phenotypic variability in Phenotypic variability in WDWD

• Age of onsetAge of onset– From two to more than 40 yearsFrom two to more than 40 years

• Homozygous for severe mutations , earlier onsetHomozygous for severe mutations , earlier onset• Milder mutations, later-onset Milder mutations, later-onset

• Presentation Presentation – Hepatic or neurological symptoms Hepatic or neurological symptoms – Most have low plasma ceruloplasmin levelsMost have low plasma ceruloplasmin levels

• Phenotypic variabilityPhenotypic variability– Over 200 mutations have been described Over 200 mutations have been described – Many WD patients are compound Many WD patients are compound

heterozygotesheterozygotes

Copper and Copper and neurodegenerative neurodegenerative diseasesdiseases• Cu-induced free radical production Cu-induced free radical production

might be involved might be involved

• Alzheimer’s disease (Alzheimer’s disease (Amyloid Precursor Amyloid Precursor

ProteinProtein) copper binding ) copper binding molecule,reduce Cu(II) to Cu(I), molecule,reduce Cu(II) to Cu(I), form reactive hydroxyl radicals form reactive hydroxyl radicals

• Creutzfeldt–Jakob (prion protein)Creutzfeldt–Jakob (prion protein)– Copper-binding protein, copper uptakeCopper-binding protein, copper uptake

Copper-binding proteinCopper-binding protein DiseaseDisease Role Role

Amyloid precursor Amyloid precursor proteinprotein

Alzheimer’s disease Alzheimer’s disease Protein involved in Protein involved in Alzheimer’s disease, is a Alzheimer’s disease, is a copper bindingcopper binding

molecule, and can reduce molecule, and can reduce Cu(II) to Cu(I), which can Cu(II) to Cu(I), which can form reactive hydroxyl form reactive hydroxyl radicals radicals

Prion proteinPrion protein Creutzfeldt–JakobCreutzfeldt–Jakob Copper uptake in Copper uptake in neurones, delivery of neurones, delivery of copper to specific target copper to specific target proteins proteins

Link between Copper and the pathogenesis of the neurodegenerative diseases

Biological Roles of Copper

Copper is an essential cofactor in many enzymes

a. Cytochrome c oxidaseb. Superoxide dismutasec. Lysyl oxidase d. Tyrosinasee. Ascorbate oxidasef. Ceruloplasmin (ferrioxidase)

Some copper enzymes in Some copper enzymes in humanshumans

Pathway of copper in the Pathway of copper in the bodybody

Concentrations above the trace essential metal

Copper ions exposed to oxidants

H2O2 inactivate CuZn-SOD(oxidation of the histidine active site)

loss of copper ion

Damage to DNAstrand breakshydroxylated base products Cancer

ROS

Genetic Diseases of Copper Metabolism

Menkes DiseaseMenkes Disease

- X-linked recessive disease.- Mainly male affected- Onset at 2-3 months of age.- Symptoms result from dysfunction of many copper-dependent enzymes due to copper deficiency.

Copper DeficiencyCopper Deficiency

• Low serum copper• Kinky hair (disulfide bonds in keratin).• connective tissue changes (Lysyl oxidase, elastin and collagen).).• Anemia (Ceruloplasmin, iron transport).).• Discoloration (tyrosinase, pigment).• Hypothermia (Cytochrome c oxidase ATP).• Skeletal deformities (Ascorbate oxidase, bone). • Heart disease (Superoxide dismutase).• Retarded growth• Metal retardation

Patient with Menkes Disease

Patient with Menkes Disease – Cont.

Biological Defect of Menkes Disease

• Defect is in small intestinal copper absorption.Defect is in small intestinal copper absorption.• The defect locus was mapped on chromosomal The defect locus was mapped on chromosomal

Xq12-13.Xq12-13.• Mutations in the Mutations in the Cu-Cu-ATP7A (MNK)ATP7A (MNK) gene are gene are

responsible for the Menkes disease.responsible for the Menkes disease.

Diagnosis and TreatmentDiagnosis and Treatment

• Family history.Family history.• Symptoms.Symptoms.• Low plasma- and serum-copper concentrations.Low plasma- and serum-copper concentrations.• Lethal. Lethal. • Most affected patients die at 2-3 years old.Most affected patients die at 2-3 years old.• Copper supplement (improve survival).Copper supplement (improve survival).• Other symptomatic and supportive treatment to Other symptomatic and supportive treatment to

improve quality of life.improve quality of life.

An allelic form of Menkes Disease

• Mutations in the Mutations in the Cu-Cu-ATP7A (MNK)ATP7A (MNK) gene.gene.• Presence of some functional ATP7A.Presence of some functional ATP7A.• Occipital horns and other bone abnormalities, Occipital horns and other bone abnormalities,

mild mental retadation, diarrhea, slight skin mild mental retadation, diarrhea, slight skin laxity, hyperextensibility, and bladder laxity, hyperextensibility, and bladder diverticula.diverticula.

• Low levels of ceruloplasmin and serum copper.Low levels of ceruloplasmin and serum copper.• Supportive treatment.Supportive treatment.

- Occipital horn syndrome or cutis laxa

Genetic Diseases of Copper Metabolism

Wilson’s DiseaseWilson’s Disease

- Autosomal recessive disease.- Onset age: 6-20 years old.- Disease is caused by overload of copper in liver.- Symptoms occurs when liver is damaged.

Symptoms of Wilson’s DiseaseSymptoms of Wilson’s Disease

• Signs of liver cirrhosis – jaundice and vomiting blood

• Kayser-Fleischer ring.

• Neurologic manifestations - speech and language problems, tremors in the arms and hands, and rigid muscles.

A. D. Patel and M. Bozdech Arch Ophthalmol. 2001;119:1556-1557

Kayser-Fleischer Ring

A. D. Patel and M. Bozdech Arch Ophthalmol. 2001;119:1556-1557

Deposition of Copper in the Basal Ganglia

Wilson Normal

Biological Defect of Wilson’s Disease

• Defect is in releasing copper into bile in liver.Defect is in releasing copper into bile in liver.• The defect locus was mapped on chromosomal The defect locus was mapped on chromosomal

13q14.3-q21.1.13q14.3-q21.1.• Wilson’s disease is caused by mutation in the Wilson’s disease is caused by mutation in the

Cu-ATP7BCu-ATP7B gene. gene.

Diagnosis and TreatmentDiagnosis and Treatment• Family history.Family history.• Symptoms.Symptoms.• High plasma-, serum-, and urine-copper High plasma-, serum-, and urine-copper

concentration.concentration.• High level of copper in the liver biopsy samples.High level of copper in the liver biopsy samples.• Low level of serum seruloplasmin.Low level of serum seruloplasmin.• Life long treatment with D-penicillamine or Life long treatment with D-penicillamine or

trientine hydrochloride.trientine hydrochloride.• Low-copper and high zinc diet.Low-copper and high zinc diet.• Patients survive if the disease detected early and Patients survive if the disease detected early and

treated correctly.treated correctly.

SummarySummary

Menkes Disease Wilson’s Disease

• X-link recessive• Defect in copper absorption in intestine• Clinic manifestation as copper defficiency• Onset early• Fetal• Caused by mutations in Cu-ATPase 7A

• Autosomal recessive• Defect in copper excretion In liver• Clinic manifestation as copper toxicity• Onset relatively late• Not fetal Caused by mutations in Cu-ATPase 7B

ZincZinc

Biological Roles of Zinc

1. As a structural component in many proteinsZinc-finger and Ring-finger proteins

2. As a catalytic component in many metallo-enzymes a. pancreatic carboxypeptidasesb. alkaline phosphatasec. various dehydrogenased. superoxide dismutase.

Genetic Diseases of Zinc Metabolism

Acrodermatitis EnterpathicaAcrodermatitis Enterpathica

- Autosomal recessive disease- Onset age: 3-10 month old infants- Symptoms occurs when breast-feeding stops.- Symptoms result from typical nutritional zinc deficiency.

Zinc DeficiencyZinc Deficiency

• Retarded growth• Skin rash and hair loss• Diarrhea and respiratory diseases• Impaired immune function• Impaired neuropsychological development

Patients with AEPatients with AE

Patients with AE – Cont.Patients with AE – Cont.

Am. J. Hum. Genet. 68:1055–1060, 2001

Biological Defect of AE

• Defect is in small intestinal zinc absorption.Defect is in small intestinal zinc absorption.

• The defect locus was mapped on chromosomal The defect locus was mapped on chromosomal 8q24.3.8q24.3.

• Mutations in the Mutations in the hZIP4hZIP4 gene are responsible for the gene are responsible for the

AE phenotype.AE phenotype. Lysosome

Secretory vesicles

Synapticvesicle

hZIP4

TGN

Transportvesicle

ER

NCis Golg

iSecretory

granule

Diagnosis and TreatmentDiagnosis and Treatment

• Family history.Family history.• Zinc deficiency symptoms.Zinc deficiency symptoms.• Low plasma- and serum-zinc Low plasma- and serum-zinc

concentrations.concentrations.• Low serum alkaline phosphatase activity.Low serum alkaline phosphatase activity.• Clinical response to oral zinc supplement.Clinical response to oral zinc supplement.• Lethal if not treated.Lethal if not treated.• Life long daily zinc supplement.Life long daily zinc supplement.

Genetic Diseases of Zinc Metabolism

Pseudo-Acrodermatitis EnterpathicaPseudo-Acrodermatitis Enterpathica

- Onset age: 3-6 month old infants- Symptoms manifest as typical nutritional zinc deficiency.- Symptoms disappear when normal diet is introduced.- Zinc content in breast milk is low than normal.

Biological defect of Pseudo-AE

• Defect is in mammary glands where zinc can not be Defect is in mammary glands where zinc can not be deposited into milk.deposited into milk.

SummarySummary

AE Disease Pseudo-AE

• Autosomal recessive• Defect in zinc absorption in intestine• Clinic manifestation as zinc deficiency• Survival with zinc supplementation• Mutations in hZIP4

• Autosomal recessive ?• Defect in zinc excretion in mammary gland• Clinic manifestation as zinc deficiency•Curable • Mutations in ZNT4 ?