2 CRYSTAL-STRUCTURES FOR CATHEPSIN-D - THE LYSOSOMAL TARGETING SIGNAL AND ACTIVE-SITE

被引:137
作者
METCALF, P
FUSEK, M
机构
[1] European Molecular Biology Lab, D6900 Heidelberg, Meyerhofstrasse 1
关键词
ASPARTIC PROTEASE; CATHEPSIN-D; INHIBITOR COMPLEX; LYSOSOMAL TARGETING; 3-DIMENSIONAL STRUCTURE;
D O I
10.1002/j.1460-2075.1993.tb05774.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Two crystal structures are described for the lysosomal aspartic protease cathepsin D (EC 3.4.23.5). The molecular replacement method was used with X-ray diffraction data to 3 angstrom resolution to produce structures for human spleen cathepsin D and for bovine liver cathepsin D complexed with the 6-peptide inhibitor pepstatin A. The lysosomal targeting region of cathepsin D defined by previous expression studies [Barnaski et al. (1990) Cell, 63, 281-219] is located in well defined electron density on the surface of the molecules. This region includes the putative binding site of the cis-Golgi phosphotransferase which is responsible for the initial sorting step for soluble proteins destined for lysosomes by phosphorylating the carbohydrates on these molecules. Carbohydrate density is visible at both expected positions on the cathepsin D molecules and, at the best defined position, four sugar residues extend towards the lysosomal targeting region. The active site of the protease and the active site cleft substrate binding subsites are described using the pepstatin inhibited structure. The model geometry for human cathepsin D has rms deviations from ideal of bonds and angles of 0.013 angstrom and 3.2-degrees respectively. For bovine cathepsin D the corresponding figures are 0.014 angstrom and 3.3-degrees. The crystallographic residuals (R factors) are 16.1% and 15.8% for the human and inhibited bovine cathepsin D models respectively. The free R factors, calculated with 10% of the data reserved for testing the models and not used for refinement, are 25.1% and 24.1% respectively.
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页码:1293 / 1302
页数:10
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