PMID: 2108713Jan 9, 1990Paper

Cloning, sequencing, expression, and site-directed mutagenesis of the gene from Clostridium perfringens encoding pyruvoyl-dependent histidine decarboxylase

Biochemistry
P D van Poelje, E E Snell

Abstract

The DNA encoding pyruvoyl-dependent histidine decarboxylase (HisDCase) of Clostridium perfringens was cloned, sequenced, and overexpressed in Escherichia coli. The gene encodes a single polypeptide of 320 amino acids, Mr 35,526, demonstrating that clostridial HisDCase, which has an (alpha beta)6 structure, is synthesized as a precursor (proHisDCase, pi 6). No pi subunits of proHisDCase were observed in crude or purified preparations of the cloned HisDCase; they appear to undergo rapid cleavage in vivo to the alpha (Mr 24,887) and beta (Mr 10,526) subunits characteristic of this HisDCase. This cleavage occurs between Ser-96 and Ser-97; Ser-97 gives rise to the catalytically essential pyruvoyl group blocking the N-termini of the alpha subunits of the active enzyme. When Ser-97 was converted to an alanyl residue by site-specific mutagenesis, the expressed, inactive protein (pi' 6) contained a single peptide species (pi', Mr 35,510) that was not cleaved either in vivo or in vitro. These results support previous conclusions that activation of the wild-type clostridial proenzyme occurs via nonhydrolytic serinolysis. Although clostridial HisDCase has only a 47% sequence similarity to HisDCase from Lactobacillus 30a, all of the residue...Continue Reading

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Citations

Jan 1, 1993·Protein Science : a Publication of the Protein Society·J ReizerM H Saier
Jul 29, 2011·Critical Reviews in Food Science and Nutrition·Daniel M LinaresMaría Fernández
Jun 29, 2002·Applied and Environmental Microbiology·Nathalie ConnilHervé Prévost
Mar 5, 2005·Applied and Environmental Microbiology·Patrick M LucasAline Lonvaud-Funel
Feb 1, 2006·Proceedings of the National Academy of Sciences of the United States of America·Jin Kwang KimKyung Hyun Kim
Mar 1, 1997·Chemistry & Biology·Y Shao, S B Kent
Mar 18, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Xin-Jun YuZhao Wang
Oct 1, 1990·Applied Biochemistry and Biotechnology·J S Dordick
Jan 7, 2011·Molecular Microbiology·Hein TripJuke S Lolkema
Mar 21, 2006·The Biochemical Journal·Yu YoshikaneToshiharu Yagi

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