Abstract:
:Bindings of cobra venom phospholipases A2 to micelles of n-hexadecylphosphorylcholine were studied by the tryptophyl fluorescence method at 25 degrees C and ionic strength 0.1. The data were analyzed by assuming that the micellar surface has multiple binding sites for the enzyme and these sites are identical and mutually independent. The enzyme binding site was found to accommodate a constant number of substrate (monomer) molecules, N = 10, 5 or 13 for N. naja atra apoenzyme and its Ca2+ complex, and N. naja kaouthia apoenzyme, respectively. The binding constant of the enzymes to the micelle, Kmic = 0.18-3.1 X 10(6) M-1, was 9-160 times greater than that to the monomeric substrate, Kmon = 2 X 10(4) M-1 (Teshima et al. (1981) J. Biochem. 89, 1163-1174). This was interpreted in terms of the presence of an additional substrate-binding site in the enzyme molecule. The binding constant of the enzyme-Ca2+ complex to the micelle was smaller than that for the apoenzyme over a wide range of pH. The pH dependence of the binding constant of the apoenzyme to the micelle was well interpreted in terms of pK shifts of two ionizable groups from 5.4 to 5.53 and 7.55 to 7.95. The pH dependence curve for the Ca2+ complex, which lacked the former transition, was interpreted in terms of the pK shift of only a single ionizable group from 7.25 to 7.55. The former ionizable group was assigned as Asp 49, to which Ca2+ can coordinate, and the latter as His 48 in the active site on the basis of the reported pK values of these ionizable groups in the apoenzyme and Ca2+ complex (Teshima et al. (1981) J. Biochem. 89, 13-20 and Teshima et al. (1982) J. Biochem. 91, 1777-1788). No participation of the alpha-amino group with a pK value of 8.55 was observed.
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Teshima K,Ikeda K,Hamaguchi K,Hayashi Kdoi
10.1093/oxfordjournals.jbchem.a134333subject
Has Abstractpub_date
1983-07-01 00:00:00pages
223-32issue
1eissn
0021-924Xissn
1756-2651journal_volume
94pub_type
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journal_title:Journal of biochemistry
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更新日期:1998-10-01 00:00:00
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更新日期:2001-07-01 00:00:00
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更新日期:2011-09-01 00:00:00
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更新日期:2004-10-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a122523
更新日期:1988-10-01 00:00:00
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更新日期:2000-05-01 00:00:00
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pub_type: 杂志文章
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更新日期:1999-06-01 00:00:00
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更新日期:2009-12-01 00:00:00
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更新日期:1990-10-01 00:00:00
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更新日期:2018-03-01 00:00:00
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更新日期:1994-07-01 00:00:00
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更新日期:1993-04-01 00:00:00
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更新日期:1982-04-01 00:00:00
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journal_title:Journal of biochemistry
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更新日期:1991-11-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1981-08-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1993-11-01 00:00:00
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pub_type: 杂志文章
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更新日期:1996-10-01 00:00:00