Zinc

History

1509, recognized as element Essentiality demonstrated

Plants: 1869 Animals: 1934

Deficiency

Considered unlikely until 1955

swine parakeratosis shown to be caused by Zn deficiency conditioned human deficiency demonstrated in 1956 1961, hypogonadal dwarfism suggested to be zinc deficiency

Facts

30th element in the periodic table (IIB element)

MW = 65.37, completely filled d orbitals

One oxidation state, namely Zn2+ Prefers tetrahedral complex formation readily complexes with amino acids, peptides, proteins and nucleotides affinity for thiols, hydroxy groups & ligands with electronrich nitrogen donors

In aqueous solutions

Not a redox active metal

Distribution

Whole body: 1.5g (female)-2.5g (male)

Skeletal Muscle Bone Skin Liver Brain Kidneys Heart Hair Blood Plasma

57% 29% 6% 5% 1.5% 0.7% 0.4% ~0.1% ~0.1%

Sources

Relatively abundant mineral Good sources: shellfish, beef and other red meats Slightly less good: Whole-grains
most in bran and germ portions 80% lost to milling phytates, hexa & penta phosphates depress absorption

P/Zn ratios of 10 or more

Relatively good sources: nuts and legumes Eggs, milk, poultry & fish diets lower than pork, beef, lamb diets High meat diets enhance absorption

280g or 10 oz fits right into food pyramid guide cys & met form stable chelate complexes

Zinc Methionine

Effect of trace mineral source on animal performance

Relative bioavailability of trace mineral sources

Whole Body Fluxes

4-15 mg/da (~0.15 mM)

Diet Zn++

Zn++ (50-100mM)
1-2 mg/da Metallothionine
Chelating Agents Phytates

Intestine

a-2 macroglobulin (30%) albumin (60%)

Pancreatic & Biliary Excretion: 4-5 mg/da

Plasma/Serum 2.4 mg

Target tissues Including Liver 1.2 g

Milk: 2-3 ug/mL Other Losses: Sweat, Skin, Hair up to 1 mg/da

Seminal Fluid: 196 ug/mL

Menstrual Loss: 0.1-0.5 mg

Feces: 3-14 mg/da

Urine: 0.4-0.6 mg/da

Dietary Factors that Affect Zn Absorption

Feed/Food source Phytate (calcium-phytatezinc complex)

Presence/Absence of other divalent cations

Fe, Ca

Mainly hexa- and pentaphosphate derivatives Highly dependent on calcium histidine, cysteine

Efficiency of absorption can vary from 15-60%

Amino Acids

Under normal conditions 1/3 of dietary Zn is absorbed Zn status alters efficiency of absorption

Uptake and retention is > in growing animals

Overview

Approximately 300 enzymes are associated with zinc Biological functions of Zn are divided into three categories

Catalytic, Structural, Regulatory

Protein synthesis Nucleic acid metabolism Carbohydrate and energy metabolism Lipid Epithelial tissue integrity Cell repair and division Vitamin A and E transport and utilization Immune function Reproductive hormones

Role in metabolism

Absorption

Absorption takes place throughout the intestine

Glycocalyx

Barrier? Storage site?

Primarily in the jejunum Some absorption in the rumen No measurable amounts absorbed from stomach cecum or colon

Absorption

In small intestine

Nonmediated (nonsaturable) process

Not affected by dietary Zn intake Stimulated by Zn depletion

Mediated (saturable) process

Absorption
Mucosa Serosa

NSBP
Zn++ Zn++

CRIP

Zn++-Albumin

Saturable = Bound to form transport ligand

Zn++

CRIP-Zn MTI-Zn MTI

Non-saturable = Passive Diffusion

Albumin
Zn++-Albumin Zn++

CRIP=cysteine-rich intestinal protein; MTI=metallothionine; NSBP, non-specfic binding protein

Transport in blood

Plasma contains approx .1% of the total zinc of the body Albumin is major portal carrier

Binds to albumin by tetrahedral ligation to sulfur atoms 70% of Zn is bound to albumin in plasma 20-30% bound to -2 macroglobulin Other plasma proteins

Transferrin, histidine-rich glycoprotein, metallothionine

Plasma Zn concns respond to external stimuli

Intake fluctuations Fasting Acute stresses

infection

Plasma Zn levels do not influence absorption from mucosa Most reductions in plasma levels reflect increased hepatic uptake

Hormonal control

Transport

Rapidly cleared from plasma by liver

Fast component of 2 pool model (T1/2 = 12.3 da)

Slow component, other tissues (T1/2 = 300 da) Bone and CNS uptake slow Pancreas, liver and kidney most rapid RBC & muscle in between Exchangeable pool & zinc status

Single dose of zinc is taken up with T1/2 = 20 s

Cellular Uptake

Hepatic uptake via a biphasic process

Contribution to overall Zn flux

Sequesters newly absorbed Zn Removes Zn from the circulation

Saturable process initial step

Temperature dependent rapid Stimulated by glucocorticoids

Linear accumulation subsequent step

slow Not affected by dietary Zn intake Does not require energy

Cellular Uptake

Erythrocytes

Depends upon bicarbonate ions

Biphasic uptake (same as liver)

Fibroblasts, proximal tubule, lymphocyte

Intracellular Transport

Zinc transporters regulate Zn ion concentrations through import, export or sequestering Zn into vesicles

Storage, toxicity

2 families exist:
ZnT- mainly exports Zn ions from cells ZIP important for Zn influx

Intracellular Transport

Number of transporters

ZnT-1: all organs, small intestine (basolateral membrane), kidney (tubular cells), placenta

Efflux Efflux & (?) intracellular vesicles Influx, intracellular retention Efflux (into milk)

ZnT-2: intestine, kidney, testis

ZnT-3: brain (synaptic vesicles) & testis

ZnT-4: mammary gland & brain

Lethal mouse transgenic

Intracellular Transport

ZIP family transporters:

Consist of:
hZIP1 hZIP2 hZIP3

Responsible for influx of Zn as well as Mn2+, Cd2+, and other divalent cations into cells

Intracellular Transport

Number of transporters

DCT1: duodenum, jejunum, kidney, bone marrow, others

Non-specific: Zn, Cd, Mn & Cu actually have slightly higher affinity than Fe, the mineral for which the transport actions of this protein was first identified. Competition between Fe & Zn & Cu

Storage

Storage sites

No specfic storage sites are recognized

Within cells, amounts sequestered within metallothionine could be considered as stores Anorexia, muscle catabolism, tissue zinc release

Metalloenzymes cling tenaciously to zinc

Serum/plasma zinc drops rapidly (~1 week) with zinc deficient diet

Zinc turnover is extensive and rapid

Two-components of turnover, fast ~12.3 days, and slow, ~300 days Usually amounts to 157-183 mg Zn

Fast pool is also called the exchangeable pool

Excretion

Lost via hair, sweat, desquamation, bile pancreatic secretions, seminal fluid, urine, feces Main endogenous loss

Secretions into gut

Bile and pancreas

Mucosal cells

Urinary and integumental losses

< 20% under normal conditions

Losses increase with trauma, muscle catabolism, and administration of chelating agents (EDTA)

Primarily in fecal material

Unabsorbed Zn Secreted Zn (endogenous sources)

From pancreatic and intestinal sources

Regulation

Metallothionein

Concentrated in liver, kidney, pancreas, intestine Acts as a Zn2+ buffer

Controls free Zn2+ level Control intracellular Zn pool responsive to both hormones and diet

Zn-binding protein, metallothionein (MT), is involved in the regulation of Zn metabolism MT is inducible by dietary Zn via the metal response element (MRE) and MTF-1 mechanism of transcriptional regulation
in cellular MT Zn binding within cells

Acute infections associated with proinflammatory cytokines increses Zn uptake into liver, bone marrow and thymus and reduces the amount going to bone, skin and intestine

Metabolic Interactions

Interactions of other divalent cations in the intestinal lumen

Fe, Sn, Cd Zn Zn Cu

Interactions

Copper High Zn diets reduce Cu absorption

electronic configuration competition

sequesters Cu in mucosal cell preventing serosal transfer

Metallothionine synthesis induced

Happens with 150mg Zn for two years Can be used with Wilsons disease patients High copper diets do not interfere with Zinc absorption

Iron

Supplements inhibit zinc absorption

Ferrous > Ferric, heme no effect Pregnant and taking >60mg Fe/day should also take Zn

Interactions

Calcium High Ca diets reduce Zn absorption

effect enhanced in phytate rich diets not sure how much of a problem in humans

post menopausal women yes, adolescent girls, no

Other
Tin (Sb), not usually high in diet, but diets high in Tin can increase fecal Zn excretion Cadmium (Cd), alter Zn distribution in body rather than altering absorption Folic acid, conjugase requires Zn

High doses sometimes impair Zn status further in low Zn situation - mechanism currently unclear

Function

Zinc-containing enzymes

More than 70 enzymes

Secondary & tertiary protein structures

Metal stabilized active sites

Examples of general types

dehydrogenases phosphatases peptidases kinases deaminases

Insulin

Function

Cu/Zn Superoxide Dismutase

General class of enzymes that protect against oxidative damage in the body.

Insulin
Zn important structurally Zn needed for insulin stored in pancreas

Functionality drops rapidly so more of a working store than a static store

Function

Nuclear transcription factors (>130)

Same protein structural role forms zinc-fingers Zn-fingers bind DNA

allow different nuclear hormones to interact with DNA via different DNA binding proteins

up to 37 fingers have been found on a single transcription factor Vit. A, Vit. D, steroid hormones, insulin-like growth factor1, growth hormone, and others bind to zinc-finger proteins to modulate gene expression

Zn is responsible for thymidine incorporation

Function

Cell Differentiation
Thymidine kinase activity Creatine kinase activity

Transcription Factors

Transcription factors
Regulate gene expression Involved in virtually all biological processes:

Development, differentiation, cell proliferation, response to external stimuli

Consists of 2 domains
DNA Binding Domain (DBD) recognizes and binds to specific DNA sequence elements in the promoter of target genes Protein-interacting Transactivation Domain (TAD) influences the rate of transcription

Zinc Finger Proteins

Zinc finger proteins are characterized by their utilization of zinc ions as structural components C2H2 zinc finger binding motif
Predominant motif in eukaryotic transcription Involved in skeletal differentiation Zinc binding motif is determined by the presence of 2 cysteine and 2 histidine residues that engage in a four coordinate bond with a singe Zn ion Bind to response elements in the upstream promoters of genes transcribed by RNA poly 2 Binds to 5S ribosomal RNA gene, and 5S RNA, and activates transcription by RNA polymerase 3.

Mech of Transcription

Function

Zinc-Finger

Function

Zinc-finger Interacting with DNA

Function

Zinc Fingers

Mutation c/ablation of binding

in case of Zif268, loss in sequence-specific DNA binding that allowed viral infection

Iron can replace Zn in fingers

Low Zn and high Fe Fe gives rise to ROS more readily

DNA damage & carcinogenesis?

Cadmium can replace Zn in fingers

Non-functional, cytotoxic

Transcription Factors

Revelation

Gene expression is controlled by specific proteins call transcription factors

Zinc containing transcription factors account for 1% of genome

Zinc plays key structural role in transcription factor proteins Ligands for transcription factors include:

Vitamin A Vitamin D Bile acids Thyroid hormones

Membrane Stability

Membrane fractions contain high concentrations of Zn

Increases rigidity of cell Competition for binding sites with redox metals

Protection from oxidative damage

Membrane Function

In deficient animals:

Failure of platelet aggregation

Due to impaired Calcium uptake Brain synaptic vesicles exhibit impaired calcium uptake Decreased plasma membrane sulfhydryl concentration

Peripheral neuropathy

Increased osmotic fragility in RBCs

Immune Function

After Zinc depletion

All functions within monocytes were impaired Cytotoxicity decreased in Natural Killer Cells Phagocytosis is reduced in neutrophils Normal function of T-cells are impaired B cells undergo apoptosis

High Zn supplementation shows alterations in cells similar to Zn depletion

Vitamin A & Zinc

Zn influences Vitamin A metabolism

Absorption, transport, and utilization

Vitamin A transport is mediated through protein synthesis

Zn deficiency can depress synthesis of retinol-binding protein in liver Retinol to retinaldehyde (retinal), for visual processes

Oxidative conversion of retinol to retinal requires Zn-dependent retinol dehydrogenase enzyme

Night Blindness

Hallmark deficiency sign for Vitamin A

Seen with Zn deficiency as well, why?

Stojanovic, Stitham and Hwa: Critical Rose of Transmembrane segment Zn binding I the structure and function of rhodopsin JBC 279(34):35932-35941, 2004

Rhodopsin proteins

Vitamin A
Zn-dependent Protein folding

Rhodopsin [11-cis-Retinal]

11-cis-Retinal

trans-Retinal + opsin

11-cis-Retinol

trans-Retinol

Zn and Vitamin A Interaction

Mechanisms of Toxicity

Excess accumulation within cells may disrupt functions of biological molecules

Protein, enzymes, DNA

Leads to toxic consequences

Anemia

Impaired copper availability

Acute excessive intakes

Local irritant to tissues and membranes

GI distress, nausea, vomiting, abdominal cramps, diarrhea

Relatively non-toxic

Sources of exposure drinking water, feed, polluted air

Deficiency

Signs

Growth retardation Delayed sexual maturation & impotence

More signs

Impaired testicular development

Hypogonadism & hypospermia Alopecia Acroorifical skin lesions

Other, glossitis, alopecia & nail dystrophy

Immune deficiencies Behavioral changes

Night blindness Impaired taste (hypoguesia) Delayed healing of wounds, burns, decubitus ulcers Impaired appetite & food intake Eye lesions including photophobia & lack of dark adaptation

Deficiency

Monogastric more susceptible

Chickens & pigs used to become deficient with high corn diets

Old enemy phytate

Ruminants resistant due to ability to break down phytates Increases urinary zinc excretion

Diabetes

Can cause deficiency

Elderly

Poor intakes & altered physiology

Deficiency During Pregnancy

Zn deficient rats failed to conceive Abnormalities of blastocyst development Offspring had high incidence of abnormalities

Deformities of brain, skull, limbs, eyes, heart, lungs

Low Zn intake during the third trimester may not have such profound effects
Main stages of differentiation are already complete Can result in low birth weight, and prolonged and difficult parturition

Deficiency During Pregnancy

Zinc Adequate

Zinc Deficient

3 days

4 days From Hurley&Schrader, 1975

Deficiency
Malformations in Zn deficiency Cleft lip Cleft palate Brain (Hydrocephalus, anencephalus or exencephalus) Micro- or agnathia Micro- or anopthalmia Clubbed feet A- or syndactyly Curly or stubby tail Dorsal herniation Heart (abnormal position) Lung (missing lobes) Urogentital
(Hydronephrosis, missing kidney, or abnormal positions)

Stress Response

Factors that decrease plasma Zn concentration

Infection Bacterial endotoxins Surgery Burns Pregnancy

IL-1 causes increased Zn uptake by liver thymus and bone marrow Severe trauma or death can result from Zn supplementation to stressed animals

2002 DRIs

Infants

UL=(x)

0-6 mo: 2 mg/d AI (4) 7mos-1 yr: 3 mg/d (5) 1-3 yrs: 3 mg/d (7) 4-8 yrs: 5 mg/d (12) 9-13 yrs: 8 mg/d (23) 14-18 yrs: (34)

Children & adolescents

Adults: 19 yrs & older (40) Men: 11 mg/da Women: 8 mg/da Pregnancy: 11-18 yrs: 12 mg/da (34) 19-50 yrs: 11 mg/day (40) Lactation: 11-18 yrs: 13mg/da (34) 19-50 yrs: 12 mg/day (40)

Males 11 mg/da Footnote Females 9 mg/da Males need more than females due to high Zn content of seminal fluids & relatively low Zn loss through menstruation