Conformational study of proteins by SAXS and EDXD: the case of trypsin and trypsinogen

被引:27
作者
Caracciolo, G
Amiconi, G
Bencivenni, L
Boumis, G
Caminiti, R
Finocchiaro, E
Maras, B
Paolinelli, C
Castellano, AC
机构
[1] Univ Roma La Sapienza, Dipartimento Fis, I-00185 Rome, Italy
[2] INFM, I-00185 Rome, Italy
[3] Univ Roma La Sapienza, Dipartimento Chim, I-00185 Rome, Italy
[4] Univ Roma La Sapienza, CNR, Ctr Mol Biol, I-00185 Rome, Italy
[5] Univ Roma La Sapienza, Dipartimento Sci Biochim, I-00185 Rome, Italy
来源
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS | 2001年 / 30卷 / 03期
关键词
beta-trypsin; trypsinogen; small angle scattering; diffraction;
D O I
10.1007/s002490000128
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
The radius of gyration (R-g) of bovine trypsinogen and beta -trypsin was measured by an energy-dispersive X-ray technique (EDXD) and by small-angle X-ray scattering (SAXS), under different solvent conditions. Both techniques gave superimposable results. The experimental evidence demonstrated that: (1) no structural modifications and/or damage occurred during the data acquisition by EDXD; (2) at pH 4 the active enzyme has one class of chloride binding sites in common with the zymogen, whereas the latter protease shows an additional class able to reverse the effects on R-g induced by chloride at low concentration; and (3) the PH profile of the R-g of both proteases does not resemble at all the pH effect on beta -trypsin activity, a result in line with the finding that the electrical potentials induced by surface charge are small in absolute magnitude and produce no gradient across the active site.
引用
收藏
页码:163 / 170
页数:8
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