Vitamin B12 and Folate Metabolism

Blood and Immunology MDP 10307 Dr Zunika Amit 20.1.2014

Objectives
List the sources of folic acid and vitamin B12 Describe the vitamins dependent enzymes Describe the relationship between folic acid and vitamin B12 Explain the absorption and transport of the vitamins Describe the diseases caused by the deficiency of these vitamins

Dietary sources of Vitamin B12

Synthesised by all types of bacteria and algae with a few exceptions. Enters human food chain by being incorporated into food of animal origin. Incorporation mainly in GIT of herbivorous. B12 are absorbed and incorporated into the tissues of the herbivorous. Thus, omnivorus and carnivorous derive B12 from the tissues or products from these animals.

Vitamin B12

Structure of Vitamin B12

Consists of cobalt called as cobamide or cobalamin cobalt is in a coordination state of six 4 coordination position occupied pyrrole rings 5th coordination position occupied by a 5,6dimethylbenzimidazole (DMB) 6th coordination position (R) by one of several different ligands; 5-deoxyadenosyl, methyl, hydroxyl (OH), cyanide (CN)

Different forms of B12

When isolated, the 6th coordination state contains cyanide called cyanocobalamin Aquacobalamin or hydroxycobalamin is the form usually found in body tissue.

Two naturally/active occurring forms of B12 : 5-deoxyadenosylcobalamin -account for 70% in the liver Methylcobalamin or methyl B12- major circulating form

Absorption, transport and cellular uptake of vitamin B12 in humans

Bhagavan, 4th edition

Absorption, transport and cellular uptake of B12 in humans

Dietary B12 released by gastric acid and bind to cobalophilins (R-protein) and intrinsic factor (IF). At acid pH, R-protein bind B12 stronger than IF (relative affinity of R-protein and IF for B12 are about 50:1) In duodenum, pancreatic proteases partially degrade R-protein allowing more of B12 to bind to IF. Transfer of B12 to IF is aided by bicarbonate (more neutral pH) of pancreatic juice .

 IF- B12 complex binds to ileal receptors on the ileal mucosa cells in the presence of Ca2+ and at neutral pH. As IF- B12 complex crosses the ileal mucosa, IF is released. B12 is transferred to plasma transport protein TCII. The TCII - B12 complex delivers B12 to all cell of the body B12 is released and TCII is degraded in the lysosome.

Vitamin B12Transport Protein

1. Cobalophilins (R-protein; R-binders), occurs in plasma, gastric juice, saliva, granulocyte and other tissues and body fluids binds one B12 molecule per cobalophilin molecule. Include transcobalamin I, II and III (TCI, TCII and TCII). TCII- principal plasma binding protein for B12 newly absorbed from the intestine. Responsible for transport of B12 to tissue Defect in B12 release from lysosome also contribute to inborn error of B12 metabolism Congenital abnormality of TCII also result in megaloblastic anemia

2. Intrinsic factor (IF) from gastric parietal cells binds one B12 molecule per IF molecule necessary for the absorption of B12 later in the terminal ileum. Lack of IF can cause poor absorption of dietary B12 Pernicious anemia Fewer rbc, rbc that are produced are abnormally large and abnormal in shape. Causes of IF deficiency; Chronic gastritis, gastrectomy, autoimmune attact

3. Internal receptor for IF-B12 complex Present on microvilli of ileal cells Defect in ileal receptor also contribute to B12 deficiency.

B12 requiring reactions

1. Conversion of methylmalonyl-CoA to succinyl-CoA -lies in the pathway for degradation of the last C3 of odd-chain fatty acids together with the degradation of certain amino acids

MethylmalonylMethylmalonyl- CoA epimerase MethylmalonylCoA mutase CoA mutase 5-deoxyadenosyl cobalamin

-5-deoxyadenosyl cobalamin is the cofactor for Methylmalonyl-CoA mutase

 Defect in activity of methylmalonyl-CoA mutase or synthesis of the deoxyadenosylcobalamin coenzyme causes L-methylmalonyl-CoA accumulation methylmalonic acid causes Severe acidosis (lowering of blood pH) Damages the CNS

Mutation in methylmalonyl-CoA mutase cause a condition called methylmalonic acidemia

2. Conversion of homocysteine to methionine B12 and folate are involved together in the conversion of homocysteine to methionine.

THF

N5-methyl-THF

homocysteine

Methionine

-methylcobalamin is , the cofactor for 5-methyltetrahydrofolatehomocysteine methyltransferase or methionine synthase (MTR)

 Absence of B12 inhibits the reaction and leads to build up of N5-methylTHF known as the THF trap. Excess homocysteine (hyperhomocysteinemia) linked to cardiovascular disease:
Elevated homocysteine levels promote oxidative damage, causing inflammation and endothelial dysfunction.

Metabolic roles of folic acid and vitamin B12 in one-carbon metabolism

Methylenetetrahydrofolate reductase (MTHFR)

MTR
B12

Neurological disorder seen in B12 deficiency?

Due to progressive demyelination of nervous tissue. Possible explanation: -Methylmalonyl-CoA compete with malonyl-CoA for fatty acid synthesis -If residual fatty acid synthesis occur, methylmalonyl-CoA substitutes for malonyl-CoA in reaction sequence. -Thus, forming branched chain fatty acid causing disruption of normal membrane structure

Folic Acid
Sources : Green leafy vegetables, fruits and liver susceptible to heat destruction

Min. daily requirements 50 g for normal, nonpregnant adult pregnant woman requires - 400 g/day

Structure of folic acid (pteroyl glutamic acid)

Folate
Folic acid has a number of derivatives known collectively as folates. Contains 3-8 (or more) glutamate residues This tail is split off to mono- (or di-) glutamate form by conjugase during absorption in the small intestine (at the brush border) 50% of the total body store is in polyglutamate form (in the liver) folic acid is reduced to active tetrahydrolate (THF) by DHF reductase form

 Circulates in plasma methyltetrahydrofolate.

primarily

as

5-

Inside cell, 5-methyl THFA may be demethylated to THFA, the active form participating in folatedependent enzymatic reactions. Monoglutamate the only form of folate that can be transported across membranes.

What does THF do?

Can accept so called C1 units from various donors and pass them on in various biosynthetic reactions. It transfers and interconverts single carbonunits at the methyl, methylene and formyl oxidation levels

What does THF do?

(continued)

Reactions with the methyl, methylene and formyl oxidation levels involves: Metabolism of certain N-containing compounds Choline, serine, glycine, methionine and histidine Biosynthesis of purine nucleotides Biosynthesis of methyl group of dTMP (the rate limiting step in DNA synthesis)

Causes of folate deficiencies:

Inadequate intake Impaired absorption (anticonvulsants and oral contraceptives) Increased demand Impaired metabolism (oral contraceptive)

Effects of folate deficiency:

Inhibition of DNA synthesis due to decreased availability of purine and dTMP Failure or slow nuclear maturation and division

Megaloblastic anemia associated with B12 deficiency is related to effect of B12 on folate metabolism.
How? When B12 is low, flux through methionine synthase step decreases: 5-methylTHF accumulates The other THF forms get depleted because reduction of 5,10-methyleneTHF to 5-MethylTHF does not stop (irreversible). Therefore, insufficient levels of the formyl and methylene derivatives for the synthesis of nucleic acid precursors.

Metabolic roles of folic acid and vitamin B12 in one-carbon metabolism

Methylenetetrahydrofolate reductase (MTHFR)

MTR
B12

Thank you
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