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
 In aqueous solutions
 One oxidation state, namely Zn2+
 Prefers tetrahedral complex formation
 Not a redox active metal
 readily complexes with amino acids, peptides, proteins and
nucleotides
 affinity for thiols, hydroxy groups & ligands with electron-
rich nitrogen donors
 Distribution
 Whole body: 1.5g (female)-2.5g (male)
 Skeletal Muscle 57%
 Bone 29%
 Skin 6%
 Liver 5%
 Brain 1.5%
 Kidneys 0.7%
 Heart 0.4%
 Hair ~0.1%
 Blood Plasma ~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
Plasma/Serum
Diet Zn++ 2.4 mg Target tissues
4-15 mg/da a-2 macroglobulin Including
(~0.15 mM) (30%) Liver
albumin
(60%) 1.2 g
Intestine
Zn++ (50-100mM) Pancreatic &
Milk: 2-3 ug/mL
1-2 mg/da Biliary Other Losses:
Metallothionine Excretion: Sweat, Skin, Hair up to
Chelating Agents 4-5 mg/da 1 mg/da
Phytates
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  Presence/Absence of
 Phytate (calcium-phytate- other divalent cations
 Fe, Ca
zinc complex)
 Mainly hexa- and
 Efficiency of absorption
pentaphosphate
can vary from 15-60%
derivatives  Under normal conditions
1/3 of dietary Zn is
 Highly dependent on absorbed
calcium  Zn status alters efficiency
 Amino Acids of absorption
 histidine, cysteine  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
 Role in metabolism
 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
 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
 Mediated (saturable) process
 Stimulated by Zn depletion
 Absorption
Mucosa Serosa
NSBP
CRIP Zn++-Albumin

Zn++ Zn++
Saturable =
Bound to CRIP-Zn Albumin
form transport MTI-Zn
ligand Zn++-Albumin
MTI
Zn++ Zn++
Non-saturable = Passive Diffusion

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 α-2 macroglobulin
 Other plasma proteins
 Transferrin, histidine-rich glycoprotein, metallothionine
 Plasma Zn concn’s 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)
 Single dose of zinc is taken up with T1/2 = 20 s
 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
 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
 Fibroblasts, proximal tubule, lymphocyte
 Biphasic uptake (same as liver)
 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
 ZnT-2: intestine, kidney, testis
 Efflux & (?) intracellular vesicles
 ZnT-3: brain (synaptic vesicles) & testis
 Influx, intracellular retention
 ZnT-4: mammary gland & brain
 Efflux (into milk)
 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
 Fast pool is also called the “exchangeable” pool
 Usually amounts to 157-183 mg Zn
 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
 Metallothionine synthesis induced
 sequesters Cu in mucosal cell preventing serosal
transfer
 Happens with 150mg Zn for two years
 Can be used with Wilson’s 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 factor-
1, 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
 Protection from oxidative damage
 Competition for binding sites with redox metals
 Membrane Function
 In deficient animals:
 Failure of platelet aggregation
 Due to impaired Calcium uptake
 Peripheral neuropathy
 Brain synaptic vesicles exhibit impaired calcium uptake
 Increased osmotic fragility in RBCs
 Decreased plasma membrane sulfhydryl concentration
 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
 Oxidative conversion of retinol to retinal requires Zn-dependent
retinol dehydrogenase enzyme
 Retinol to retinaldehyde (retinal), for visual processes
 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  More signs
 Growth retardation
 Night blindness
 Delayed sexual maturation &
impotence  Impaired taste (hypoguesia)
 Impaired testicular development  Delayed healing of wounds,
 Hypogonadism & hypospermia burns, decubitus ulcers
 Alopecia  Impaired appetite & food
 Acroorifical skin lesions intake
 Other, glossitis, alopecia & nail  Eye lesions including
dystrophy photophobia & lack of dark
 Immune deficiencies adaptation
 Behavioral changes
 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
 Diabetes
 Increases urinary zinc excretion
 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
Zinc
Adequate
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 DRI’s

 Infants UL=(x) Adults: 19 yrs & older (40)

Men: 11 mg/da
 0-6 mo: 2 mg/d AI (4) Women: 8 mg/da
 Children & adolescents
Pregnancy:
 7mos-1 yr: 3 mg/d (5) 11-18 yrs: 12 mg/da (34)
 1-3 yrs: 3 mg/d (7) 19-50 yrs: 11 mg/day (40)
 4-8 yrs: 5 mg/d (12) Lactation:
11-18 yrs: 13mg/da (34)
 9-13 yrs: 8 mg/d(23)
19-50 yrs: 12 mg/day (40)
 14-18 yrs: (34)
 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