Prime region subsite specificity characterization of human cathepsin D: The dominant role of position 128

被引:13
作者
Beyer, BM [1 ]
Dunn, BM [1 ]
机构
[1] Univ Florida, Coll Med, Dept Biochem & Mol Biol, Gainesville, FL 32610 USA
关键词
aspartic proteinase; cathepsin D; chromogenic substrate; prime region subsites; short" pseudocathepsin D; substrate specificity;
D O I
10.1002/pro.5560070109
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
In order to contribute to our understanding of cathepsin D (CatD) active site specificity, two series of chromogenic octapeptides with systematic substitutions in positions P-2' and P-3' were synthesized. This panel was characterized with native human liver cathepsin D (nHuCatD) and yielded information concerning specificity trends within the S-2' and S-3' subsites. The pepstatin inhibited crystal structure of nHuCatD (Baldwin et al., 1993) was then utilized in conjunction with these subsite preference data to identify residues suspected of contributing to "prime" side subsite specificity. These residues were targeted for site-directed mutagenesis using the re-engineered recombinant model, "short" pseudocathepsin D (Beyer & Dunn, 1996). As a result of these analyses it was determined that prime region subsites do contribute to the unique specificity of human CatD. Furthermore, it was ascertained that the poly-proline loop does not have an active role in S-3' subsite specificity. Lastly, it appears that Ile128 has a dominant role on S-2' subsite specificity whereas Val130 does not.
引用
收藏
页码:88 / 95
页数:8
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