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Purification and characterization of the proteinase ECP 32 from Escherichia coli A2 strain

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dc.contributor.author Matveyev V.
dc.contributor.author Usmanova A.
dc.contributor.author Morozova A.
dc.contributor.author Collins J.
dc.contributor.author Khaitlina S.
dc.date.accessioned 2018-09-17T21:42:06Z
dc.date.available 2018-09-17T21:42:06Z
dc.date.issued 1996
dc.identifier.issn 0167-4838
dc.identifier.uri https://dspace.kpfu.ru/xmlui/handle/net/135323
dc.description.abstract The proteinase previously described as an unidentified component of E. coli A2 extracts which hydrolyses actin at a new cleavage site (Khaitlina et al. (1991) FEBS Lett. 279, 49) was isolated and further characterized. A chromatographic method of proteinase purification was developed by which a purity of more than 80% was attained. The enzyme was identified as a single, 32 kDa polypeptide (ECP 32) by SDS-PAGE and non-denaturing electrophoresis as well as by ion-exchange chromatography and gel filtration. The N-terminal sequence of ECP 32 was determined to be: AKTSSAGVVIRDIFL. The activity of ECP 32 is inhibited by o-phenanthroline, EDTA, EGTA and zincone. The EDTA-inactivated enzyme can be reactivated by cobalt, nickel and zinc ions. Based on these properties ECP 32 was classified as a metalloproteinase (EC 3.4.24). Limited proteolysis of skeletal muscle actin between Gly-42 and Val-43 was observed at enzyme substrate mass ratios of 1:25 to 1:3000. Two more sites between Ala-29 and Val-30, and between Ser-33 and Ile-34 were cleaved by ECP 32 in heat- or EDTA-inactivated actin. Besides actin, only histones and DNA-binding protein HU were found to be substrates of the proteinase, confirming its high substrate specificity. Its molecular mass, N-terminal sequence and enzymatic properties distinguish ECP 32 from any known metalloproteinases of E. coli, and we therefore conclude that it is a new enzyme.
dc.relation.ispartofseries Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
dc.subject Characterization
dc.subject E. coli
dc.subject Gel filtration (size exclusion)
dc.subject Ion-exchange chromatography
dc.subject Metalloproteinase
dc.subject Proteinae ECP 32
dc.subject Purification
dc.title Purification and characterization of the proteinase ECP 32 from Escherichia coli A2 strain
dc.type Article
dc.relation.ispartofseries-issue 1
dc.relation.ispartofseries-volume 1296
dc.collection Публикации сотрудников КФУ
dc.relation.startpage 55
dc.source.id SCOPUS01674838-1996-1296-1-SID0002013019


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  • Публикации сотрудников КФУ Scopus [24551]
    Коллекция содержит публикации сотрудников Казанского федерального (до 2010 года Казанского государственного) университета, проиндексированные в БД Scopus, начиная с 1970г.

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