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. 1977 Dec;60(6):1381–1392. doi: 10.1172/JCI108899

Absorption, Plasma Transport, and Cellular Retention of Cobalamin Analogues in the Rabbit

EVIDENCE FOR THE EXISTENCE OF MULTIPLE MECHANISMS THAT PREVENT THE ABSORPTION AND TISSUE DISSEMINATION OF NATURALLY OCCURRING COBALAMIN ANALOGUES

J Fred Kolhouse 1, Robert H Allen 1
PMCID: PMC372496  PMID: 915005

Abstract

Analogues of cobalamin (Cbl; vitamin B12) are prevalent in nature as a result of bacterial synthesis, and are of additional interest because of their potential use as antimetabolites and chemotherapeutic agents. We have synthesized 14 Cbl analogues containing 57Co and have compared their gastrointestinal absorption, plasma transport, and cellular retention to that of [58Co]Cbl in rabbits.

Many of the Cbl analogues were bound with low affinity by intrinsic factor, and none of these [57Co]Cbl analogues were taken up by the ileum or absorbed into the body in amounts comparable to that of [58Co]Cbl. The Cbl analogues that were bound by intrinsic factor with high affinity were taken up by the ileum but, in many cases, they were retained there in significant amounts.

Most of the Cbl analogues were bound by plasma transcobalamin II with high affinity and all of these transcobalamin II-[57Co]Cbl analogue complexes were taken up by a variety of tissues in a manner that was indistinguishable from that of transcobalamin II-[58Co]Cbl. The few analogues that were bound by transcobalamin II with low affinity were taken up by tissues in lesser amounts, and 20-70% of these analogues was rapidly excreted in the urine as occurs with native Cbl when it is present in plasma in unbound form.

All of the Cbl analogues were bound by the granulocyte R-type Cbl-binding protein with high affinity and all of the R-type protein-[57Co]Cbl analogue complexes were cleared rapidly from plasma exclusively by hepatocytes as occurs with R-type protein-[58Co]Cbl. Some Cbl analogues were released back into the plasma and were disseminated among a variety of tissues via transcobalamin II as occurs with native Cbl. Other Cbl analogues were retained in the liver and eventually excreted in the feces and urine without accumulating in other tissues.

These studies indicate that intrinsic factor and the ileum prevent certain Cbl analogues from entering the body and that the granulocyte R-type protein and hepatocytes prevent the dissemination of certain Cbl analogues that may gain entry such as during infections with Cbl analogue-producing bacteria. The fact that transcobalamin II binds and transports a large number of Cbl analogues indicates that these protective mechanisms can be circumvented and supports the feasibility of using Cbl analogues as antimetabolites in vivo.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen R. H. Human vitamin B12 transport proteins. Prog Hematol. 1975;9:57–84. [PubMed] [Google Scholar]
  2. Allen R. H., Majerus P. W. Isolation of vitamin B12-binding proteins using affinity chromatography. I. Preparation and properties of vitamin B12-sepharose. J Biol Chem. 1972 Dec 10;247(23):7695–7701. [PubMed] [Google Scholar]
  3. Allen R. H., Majerus P. W. Isolation of vitamin B12-binding proteins using affinity chromatography. II. Purification and properties of a human granulocyte vitamine B12-binding protein. J Biol Chem. 1972 Dec 10;247(23):7702–7708. [PubMed] [Google Scholar]
  4. Allen R. H., Mehlman C. S. Isolation of gastric vitamin B 12 -binding proteins using affinity chromatography. I. Purification and properties of human intrinsic factor. J Biol Chem. 1973 May 25;248(10):3660–3669. [PubMed] [Google Scholar]
  5. Ashwell G., Morell A. G. The role of surface carbohydrates in the hepatic recognition and transport of circulating glycoproteins. Adv Enzymol Relat Areas Mol Biol. 1974;41(0):99–128. doi: 10.1002/9780470122860.ch3. [DOI] [PubMed] [Google Scholar]
  6. BARKER H. A., SMYTH R. D., WEISSBACH H., TOOHEY J. I., LADD J. N., VOLCANI B. E. Isolation and properties of crystalline cobamide coenzymes containing benzimidazole or 5, 6-dimethylbenzimidazole. J Biol Chem. 1960 Feb;235:480–488. [PubMed] [Google Scholar]
  7. BUNGE M. B., SCHILLING R. F., SCHLOESSER L. L. Intrinsic factor studies. IV. Selective absorption and binding of cyanocobalamin by gastric juice in the presence of excess pseudovitamin B12 or 5, 6-dimethylbenzimidazole. J Lab Clin Med. 1956 Nov;48(5):735–744. [PubMed] [Google Scholar]
  8. Barker H. A. Corrinoid-dependent enzymic reactions. Annu Rev Biochem. 1972;41:55–90. doi: 10.1146/annurev.bi.41.070172.000415. [DOI] [PubMed] [Google Scholar]
  9. Burger R. L., Mehlman C. S., Allen R. H. Human plasma R-type vitamin B12-binding proteins. I. Isolation and characterization of transcobalamin I. TRANSCOBALAMIN III. and the normal granulocyte vitamin B12-binding protein. J Biol Chem. 1975 Oct 10;250(19):7700–7706. [PubMed] [Google Scholar]
  10. Burger R. L., Schneider R. J., Mehlman C. S., Allen R. H. Human plasma R-type vitamin B12-binding proteins. II. The role of transcobalamin I, transcobalamin III, and the normal granulocyte vitamin B12-binding protein in the plasma transport of vitamin B12. J Biol Chem. 1975 Oct 10;250(19):7707–7713. [PubMed] [Google Scholar]
  11. COATES M. E., DAVIES M. K., HARRISON G. F. Antivitamin B12 activity for the growing chick and developing chick embryo of some analogs of cyanocobalamin. Arch Biochem Biophys. 1960 Mar;87:93–99. doi: 10.1016/0003-9861(60)90128-4. [DOI] [PubMed] [Google Scholar]
  12. Carmel R., Herbert V. Deficiency of vitamin B12-binding alpha globulin in two brothers. Blood. 1969 Jan;33(1):1–12. [PubMed] [Google Scholar]
  13. Corcino J., Krauss S., Waxman S., Herbert V. Release of vitamin B12--binding protein by human leukocytes in vitro. J Clin Invest. 1970 Dec;49(12):2250–2255. doi: 10.1172/JCI106444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Friedman P. A., Shia M. A., Wallace J. K. A saturable high affinity binding site for transcobalamin II-vitamin B12 complexes in human placental membrane preparations. J Clin Invest. 1977 Jan;59(1):51–58. doi: 10.1172/JCI108621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. GOTTLIEBLAU K. S., WASSERMAN L. R., HERBERT V. RAPID CHARCOAL ASSAY FOR INTRINSIC FACTOR (IF), GASTRIC JUICE UNSATURATED B12 BINDING CAPACITY, ANTIBODY TO IF, AND SERUM UNSATURATED B12 BINDING CAPACITY. Blood. 1965 Jun;25:875–884. [PubMed] [Google Scholar]
  16. Godfrey J. M., Bonnett R., Math V. B. Cyano-13-epicobalamin(neovitamin B 12 )and its relatives. J Chem Soc Perkin 1. 1971;22:3736–3743. doi: 10.1039/j39710003736. [DOI] [PubMed] [Google Scholar]
  17. Gottlieb C. W., Retief F. P., Herbert V. Blockade of vitamin B12-binding sites in gastric juice, serum and saliva by analogues and derivatives of vitamin B12 and by antibody to intrinsic factor. Biochim Biophys Acta. 1967 Aug 29;141(3):560–572. doi: 10.1016/0304-4165(67)90185-7. [DOI] [PubMed] [Google Scholar]
  18. Halpern R. M., Halpern B. C., Clark B. R., Ashe H., Hardy D. N., Jenkinson P. Y., Chou S. C., Smith R. A. New approach to antifolate treatment of certain cancers as demonstrated in tissue culture. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4018–4022. doi: 10.1073/pnas.72.10.4018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hippe E., Haber E., Olesen H. Nature of vitamin B 12 binding. II. Steric orientation of vitamin B 12 on binding and number of combining sites of human intrinsic factor and the transcobalamins. Biochim Biophys Acta. 1971 Jul 25;243(1):75–82. [PubMed] [Google Scholar]
  20. Hippe E., Olesen H. Nature of vitamin B 12 binding. 3. Thermodynamics of binding to human intrinsic factor and transcobalamins. Biochim Biophys Acta. 1971 Jul 25;243(1):83–88. [PubMed] [Google Scholar]
  21. Hoffman R. M., Erbe R. W. High in vivo rates of methionine biosynthesis in transformed human and malignant rat cells auxotrophic for methionine. Proc Natl Acad Sci U S A. 1976 May;73(5):1523–1527. doi: 10.1073/pnas.73.5.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hooper D. C., Alpers D. H., Burger R. L., Mehlman C. S., Allen R. H. Characterization of ileal vitamin B12 Binding using homogeneous human and hog intrinsic factors. J Clin Invest. 1973 Dec;52(12):3074–3083. doi: 10.1172/JCI107506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kane S. P., Peters T. J. Analytical subcellular fractionation of human granulocytes with reference to the localization of vitamin B12-binding proteins. Clin Sci Mol Med. 1975 Aug;49(2):171–182. doi: 10.1042/cs0490171. [DOI] [PubMed] [Google Scholar]
  24. Kolhouse J. F., Allen R. H. Recognition of two intracellular cobalamin binding proteins and their identification as methylmalonyl-CoA mutase and methionine synthetase. Proc Natl Acad Sci U S A. 1977 Mar;74(3):921–925. doi: 10.1073/pnas.74.3.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lengyel P., Mazumder R., Ochoa S. MAMMALIAN METHYLMALONYL ISOMERASE AND VITAMIN B(12) COENZYMES. Proc Natl Acad Sci U S A. 1960 Oct;46(10):1312–1318. doi: 10.1073/pnas.46.10.1312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Masson P. L., Heremans J. F., Schonne E. Lactoferrin, an iron-binding protein in neutrophilic leukocytes. J Exp Med. 1969 Sep 1;130(3):643–658. doi: 10.1084/jem.130.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mathan V. I., Babior B. M., Donaldson R. M., Jr Kinetics of the attachment of intrinsic factor-bound cobamides to ileal receptors. J Clin Invest. 1974 Sep;54(3):598–608. doi: 10.1172/JCI107797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mellman I. S., Youngdahl-Turner P., Willard H. F., Rosenberg L. E. Intracellular binding of radioactive hydroxocobalamin to cobalamin-dependent apoenzymes in rat liver. Proc Natl Acad Sci U S A. 1977 Mar;74(3):916–920. doi: 10.1073/pnas.74.3.916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. PERLAMN D., BARRETT J. B. Biosynthesis of cobalamin analogues by propionibacterium arabinosum. Can J Microbiol. 1958 Feb;4(1):9–15. doi: 10.1139/m58-002. [DOI] [PubMed] [Google Scholar]
  30. Pletsch Q. A., Coffey J. W. Properties of the proteins that bind vitamin B 12 in subcellular fractions of rat liver. Arch Biochem Biophys. 1972 Jul;151(1):157–167. doi: 10.1016/0003-9861(72)90484-5. [DOI] [PubMed] [Google Scholar]
  31. Rosenberg L. E., Patel L., Lilljeqvist A. C. Absence of an intracellular cobalamin-binding protein in cultured fibroblasts from patients with defective synthesis of 5'-deoxyadenosylcobalamin and methylcobalamin. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4617–4621. doi: 10.1073/pnas.72.11.4617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schneider R. J., Burger R. L., Mehlman C. S., Allen R. H. The role and fate of rabbit and human transcobalamin II in the plasma transport of vitamin B12 in the rabbit. J Clin Invest. 1976 Jan;57(1):27–38. doi: 10.1172/JCI108265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Scott J. M., Bloomfield F. J., Stebbins R., Herbert V. Studies on derivation of transcobalamin 3 from granulocytes. Enhancement by lithium and elimination by fluoride of in vitro increments in vitamin B12-binding capacity. J Clin Invest. 1974 Jan;53(1):228–239. doi: 10.1172/JCI107543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Siddons R. C., Spence J. A., Dayan A. D. Experimental vitamin B12 deficiency in the baboon. Adv Neurol. 1975;10:239–252. [PubMed] [Google Scholar]
  35. Stenman U. H. Intrinsic factor and the vitamin B12 binding proteins. Clin Haematol. 1976 Oct;5(3):473–495. [PubMed] [Google Scholar]
  36. Yamada R. H., Hogenkamp H. P. The synthesis of a 5'-deoxyadenosylcobalamin-agarose adsorbent and its utility in the purification of ribonucleotide reductase. J Biol Chem. 1972 Oct 10;247(19):6266–6270. [PubMed] [Google Scholar]

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