Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1976 Jun 1;155(3):615–621. doi: 10.1042/bj1550615

The determination of methionine in proteins by gas-liquid chromatography.

G M Ellinger, A Duncan
PMCID: PMC1172884  PMID: 949322

Abstract

Intact methionine residues in proteins were rapidly and precisely determined by measuring methyl thiocyanate released during the reaction with CNBr and separated by g.l.c. Conditions for the reaction and for chromatography on columns of Porapak P-S are described. The recovery of methyl thiocyanate from several methionine derivatives and analogues were examined. Carbamoylmethionine was adopted as a stable primary standard and ethyl thiocyanate as internal standard. The measured methionine content of several isolated proteins was close to the theoretical value indicated by previous work and the results for these and a range of food proteins agreed well with results obtained by ion-exchange chromatography after performic acid oxidation. Since CNBr does not react with methionine sulphoxide and a preliminary hydrolysis is not required, the method discriminates between methionine and any methionine sulphoxide that may be present. It could be useful in studies on the nutritional availability of methionine in processed foods.

Full text

PDF
615

Selected References

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

  1. Braunitzer G., Chen R., Schrank B., Stangl A. Die Sequenzanalyse des beta-Lactoglobulins. Hoppe Seylers Z Physiol Chem. 1973 Aug;354(8):867–878. [PubMed] [Google Scholar]
  2. CANFIELD R. E. THE AMINO ACID SEQUENCE OF EGG WHITE LYSOZYME. J Biol Chem. 1963 Aug;238:2698–2707. [PubMed] [Google Scholar]
  3. CARPENTER K. J., ELLINGER G. M., MUNRO M. I., ROLFE E. J. Fish products as protein supplements to cereals. Br J Nutr. 1957;11(2):162–173. doi: 10.1079/bjn19570033. [DOI] [PubMed] [Google Scholar]
  4. Clark B. R., Ashe H., Halpern R. M., Smith R. A. A method for determination of methionine containing radioactivity in the thiomethyl moiety. Anal Biochem. 1974 Sep;61(1):243–247. doi: 10.1016/0003-2697(74)90351-0. [DOI] [PubMed] [Google Scholar]
  5. DEDMAN M. L., FARMER T. H., MORRIS C. J. Studies on pituitary adrenocorticotrophin. 3. Identification of the oxidation-reduction centre. Biochem J. 1961 Feb;78:348–352. doi: 10.1042/bj0780348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ellis K. J. Errors inherent in the use of piston activated pipettes. Anal Biochem. 1973 Oct;55(2):609–614. doi: 10.1016/0003-2697(73)90150-4. [DOI] [PubMed] [Google Scholar]
  7. FORD J. E. A microbiological method for assessing the nutritional value of proteins. 2. The measurement of 'available' methionine, leucine, isoleucine, arginine, histidine, tryptophan and valine. Br J Nutr. 1962;16:409–425. doi: 10.1079/bjn19620042. [DOI] [PubMed] [Google Scholar]
  8. GROSS E., WITKOP B. Nonenzymatic cleavage of peptide bonds: the methionine residues in bovine pancreatic ribonuclease. J Biol Chem. 1962 Jun;237:1856–1860. [PubMed] [Google Scholar]
  9. HIRS C. H., MOORE S., STEIN W. H. Peptides obtained by tryptic hydrolysis of performic acid-oxidized ribonuclease. J Biol Chem. 1956 Apr;219(2):623–642. [PubMed] [Google Scholar]
  10. Hofmann K., Visser J. P., Finn F. M. Studies on polypeptides. 43. Synthesis of S-peptide 1-20 by two routes. J Am Chem Soc. 1969 Aug 13;91(17):4883–4887. doi: 10.1021/ja01045a050. [DOI] [PubMed] [Google Scholar]
  11. Inglis A. S., Edman P. Mechanism of cyanogen bromide reaction with methionine in peptides and proteins. I. Formation of imidate and methyl thiocyanate. Anal Biochem. 1970 Sep;37(1):73–80. doi: 10.1016/0003-2697(70)90259-9. [DOI] [PubMed] [Google Scholar]
  12. Jori G., Galiazzo G., Marzotto A., Scoffone E. Selective and reversibe photo-oxidation of the methionyl residues in lysozyme. J Biol Chem. 1968 Aug 25;243(16):4272–4278. [PubMed] [Google Scholar]
  13. King T. P., Spencer M. Structural studies and organic ligand-binding properties of bovine plasma albumin. J Biol Chem. 1970 Nov 25;245(22):6134–6148. [PubMed] [Google Scholar]
  14. LEA C. H., HANNAN R. S. Studies of the reaction between proteins and reducing sugars in the 'dry' state. III. Nature of the protein groups reacting. Biochim Biophys Acta. 1950 Jun;5(3/4):433–454. doi: 10.1016/0006-3002(50)90189-2. [DOI] [PubMed] [Google Scholar]
  15. Link T. P., Stark G. R. S-methylmethionine-29 ribonuclease A. I. Preparation and proof of structure. J Biol Chem. 1968 Mar 25;243(6):1082–1088. [PubMed] [Google Scholar]
  16. Miller E. L., Carpenter K. J., Milner C. K. Availability of sulphur amino acids in protein foods. 3. Chemical and nutritional changes in heated cod muscle. Br J Nutr. 1965;19(4):547–564. doi: 10.1079/bjn19650049. [DOI] [PubMed] [Google Scholar]
  17. Miller S. A., Tannenbaum S. R., Seitz A. W. Utilization of L-methionine sulfoxide by the rat. J Nutr. 1970 Aug;100(8):909–915. doi: 10.1093/jn/100.8.909. [DOI] [PubMed] [Google Scholar]
  18. NEUMANN N. P., MOORE S., STEIN W. H. Modification of the methionine residues in ribonuclease. Biochemistry. 1962 Jan;1:68–75. doi: 10.1021/bi00907a011. [DOI] [PubMed] [Google Scholar]
  19. Pieniaźek D., Rakowska M., Szkilladziowa W., Grabarek Z. Estimation of available methionine and cysteine in proteins of food products by in vivo and in vitro methods. Br J Nutr. 1975 Sep;34(2):175–190. doi: 10.1017/s0007114575000232. [DOI] [PubMed] [Google Scholar]
  20. RAY W. J., Jr, KOSHLAND D. E., Jr Identification of amino acids involved in phosphoglucomutase action. J Biol Chem. 1962 Aug;237:2493–2505. [PubMed] [Google Scholar]
  21. Rajagopalan T. G., Moore S., Stein W. H. Pepsin from pepsinogen. Preparation and properties. J Biol Chem. 1966 Nov 10;241(21):4940–4950. [PubMed] [Google Scholar]
  22. Schroeder W. A., Shelton J. B., Shelton J. R. An examination of conditions for the cleavage of polypeptide chains with cyanogen bromide: application to catalase. Arch Biochem Biophys. 1969 Mar;130(1):551–556. doi: 10.1016/0003-9861(69)90069-1. [DOI] [PubMed] [Google Scholar]
  23. Shelley H. J. Carbohydrate metabolism in the foetus and the newly born. Proc Nutr Soc. 1969 Mar;28(1):42–49. doi: 10.1079/pns19690008. [DOI] [PubMed] [Google Scholar]
  24. Smith M. B., Back J. F. Studies on ovalbumin. V. The amino acid composition and some properties of chicken, duck, ad turkey ovalbumins. Aust J Biol Sci. 1970 Dec;23(6):1221–1227. doi: 10.1071/bi9701221. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES