Abstract:
:Carboxypeptidase A-catalyzed hydrolysis of peptides and depsipeptides is competitively inhibited by N-(1-carboxy-5-t-butyloxycarbonylaminopentyl)-L-phenylalanine (Boc-CA-Phe, Ki = 1.3 microM) and the angiotensin converting enzyme inhibitor, N-(1-carboxy-5-carbobenzoxyaminopentyl)-glycyl-L-phenylalanine (Z-CA-Gly-Phe, Ki = 4.5 microM). The latter compound is actually a slow substrate of carboxypeptidase. Indirect observation of inhibitor binding by stopped-flow measurement of radiationless energy transfer between carboxypeptidase tryptophans and dansylated substrates reveals slow binding for both compounds. The visible absorption spectrum of the complex of cobalt(II)-substituted carboxypeptidase and Z-CA-Gly-Phe, which differs from the corresponding spectrum of the Boc-CA-Phe complex, is remarkable in its resemblance to the spectrum of the complex between Co(II)carboxypeptidase and a transient intermediate previously observed during hydrolysis of peptide substrates. The spectrum slowly changes to that of the free enzyme indicating hydrolysis. Chromatographic quantitation of substrate and products confirms that carboxypeptidase converts Z-CA-Gly-Phe to Z-CA-Gly and L-Phe with an apparent kcat of 0.02 s-1. Absorption spectroscopy indicates that the Z-CA-Gly-Phe-Co(II)carboxypeptidase spectrum is not that of bound products. Moreover, spectral titrations indicate that the products (both with spectral Ki values of about 3 mM), as well as D-Phe, compete for the same site on the enzyme.
journal_name
J Inorg Biochemjournal_title
Journal of inorganic biochemistryauthors
Martin MT,Holmquist B,Riordan JFdoi
10.1016/0162-0134(89)80011-xsubject
Has Abstractpub_date
1989-05-01 00:00:00pages
39-50issue
1eissn
0162-0134issn
1873-3344pii
0162-0134(89)80011-Xjournal_volume
36pub_type
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journal_title:Journal of inorganic biochemistry
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journal_title:Journal of inorganic biochemistry
pub_type: 杂志文章
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journal_title:Journal of inorganic biochemistry
pub_type: 杂志文章
doi:10.1016/j.jinorgbio.2005.01.017
更新日期:2005-05-01 00:00:00
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journal_title:Journal of inorganic biochemistry
pub_type: 杂志文章
doi:10.1016/j.jinorgbio.2011.01.009
更新日期:2011-05-01 00:00:00
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pub_type: 杂志文章
doi:10.1016/s0162-0134(03)00129-6
更新日期:2003-07-01 00:00:00