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. 1983 Feb;80(4):973–977. doi: 10.1073/pnas.80.4.973

Conversion of prohistidine decarboxylase to histidine decarboxylase: Peptide chain cleavage by nonhydrolytic serinolysis

Paul A Recsei *, Quang K Huynh *, Esmond E Snell *,†
PMCID: PMC393510  PMID: 6405382

Abstract

Unlabeled prohistidine decarboxylase and prohistidine decarboxylase containing L-[carboxyl-18O]serine or L-[hydroxyl-18O]serine were isolated in homogeneous form from mutant 3 of Lactobacillus 30a grown with the appropriately labeled serine. There was no randomization or redistribution of label during growth, isolation of the protein, or enzymatic hydrolysis and reisolation of the labeled amino acids. These proteins were used to show that during proenzyme activation, in which individual π subunits of the proenzyme are converted to α and β subunits of the active enzyme [Formula: see text] (in which π, α, and β subunits have the partial structures shown and Prv designates a pyruvoyl group), no 18O from H218O is incorporated into the newly formed carboxyl terminus (Ser-81) of the β chain, although no labilization of 18O from proenzyme labeled with L-[carboxyl-18O]serine occurred when the proenzyme was activated in H216O by the same procedures. The additional oxygen atom present in the carboxyl group of Ser-81 of the β subunit is transferred from the hydroxyl group of Ser-82 of the proenzyme during the activation reaction. The same result was obtained with wild-type enzyme formed intracellularly. Peptide bond cleavage during activation of the proenzyme thus proceeds by a hitherto unobserved direct or indirect “serinolysis” coupled to α,β-elimination at Ser-82 to yield the pyruvoyl group of the α subunit, rather than by hydrolysis. Possible mechanisms for the reaction are discussed briefly.

Keywords: acyl transfer; α,β-elimination; pyruvoyl proteins; 18O-labeled proteins; [18O]serine

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Selected References

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

  1. Bergström K., Gürtler J. Trimethylsilylation of amino acids. II. Gas chromatographic and structural studies on trimethylsilyl derivatives of straight chain amino acids. Acta Chem Scand. 1970;25(1):175–188. doi: 10.3891/acta.chem.scand.25-0175. [DOI] [PubMed] [Google Scholar]
  2. GUIRARD B. M., SNELL E. E. NUTRITIONAL REQUIREMENTS OF LACTOBACILLUS 30A FOR GROWTH AND HISTIDINE DECARBOXYLASE PRODUCTION. J Bacteriol. 1964 Feb;87:370–376. doi: 10.1128/jb.87.2.370-376.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  4. Murphy R. C., Clay K. L. Synthesis and back exchange of 18O labeled amino acids for use as internal standards with mass spectrometry. Biomed Mass Spectrom. 1979 Jul;6(7):309–314. doi: 10.1002/bms.1200060709. [DOI] [PubMed] [Google Scholar]
  5. Recsei P. A., Snell E. E. Histidine decarboxylase of Lactobacillus 30a. Comparative properties of wild type and mutant proenzymes and their derived enzymes. J Biol Chem. 1982 Jun 25;257(12):7196–7202. [PubMed] [Google Scholar]
  6. Recsei P. A., Snell E. E. Prohistidine decarboxylase from Lactobacillus 30a. A new type of zymogen. Biochemistry. 1973 Jan 30;12(3):365–371. doi: 10.1021/bi00727a001. [DOI] [PubMed] [Google Scholar]
  7. Riley W. D., Snell E. E. Histidine decarboxylase of Lactobacillus 30a. IV. The presence of covalently bound pyruvate as the prosthetic group. Biochemistry. 1968 Oct;7(10):3520–3528. doi: 10.1021/bi00850a029. [DOI] [PubMed] [Google Scholar]
  8. Riley W. D., Snell E. E. Histidine decarboxylase of Lactobacillus 30a. V. Origin of enzyme-bound pyruvate and separation of nonidentical subunits. Biochemistry. 1970 Mar 31;9(7):1485–1491. doi: 10.1021/bi00809a002. [DOI] [PubMed] [Google Scholar]
  9. Vaaler G. L., Recsei P. A., Fox J. L., Snell E. E. Histidine decarboxylase of Lactobacillus 30a. Comparative sequences of the beta chain from wild type and mutant enzymes. J Biol Chem. 1982 Nov 10;257(21):12770–12774. [PubMed] [Google Scholar]
  10. Vandenheuvel W. J., Cohen J. S. Gas-liquid chromatography-mass spectrometry of carbon-13 enriched amino acids as trimethylsilyl derivatives. Biochim Biophys Acta. 1970 May 12;208(2):251–259. doi: 10.1016/0304-4165(70)90243-6. [DOI] [PubMed] [Google Scholar]
  11. Weiner H., White W. N., Hoare D. G., Koshland D. E., Jr The formation of anhydrochymotrypsin by removing the elements of water from the serine at the active site. J Am Chem Soc. 1966 Aug 20;88(16):3851–3859. doi: 10.1021/ja00968a033. [DOI] [PubMed] [Google Scholar]

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