HUMAN LIVER CATHEPSIN-D - PURIFICATION, CRYSTALLIZATION AND PRELIMINARY-X-RAY DIFFRACTION ANALYSIS OF A LYSOSOMAL-ENZYME

被引：25

作者：

GULNIK, S

BALDWIN, ET

TARASOVA, N

ERICKSON, J

机构：

[1] NCI,FCRDC,PRI DYNCORP,CTR BIOMED SUPERCOMP,STRUCT BIOCHEM PROGRAM,FREDERICK,MD 21702

[2] NCI,FCRDC,ABL BRP,MACROMOLEC STRUCT LAB,FREDERICK,MD 21702

来源：

JOURNAL OF MOLECULAR BIOLOGY | 1992年 / 227卷 / 01期

关键词：

CATHEPSIN-D; ASPARTIC PROTEINASE; LYSOSOMAL ENZYME; PURIFICATION; CRYSTALLIZATION;

D O I：

10.1016/0022-2836(92)90696-H

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：

The two-chain form of active cathepsin D, a glycosylated, lysosomal aspartic proteinase, has been isolated from human liver. Isoelectric focusing revealed two major species of enzyme that differed by approximately 0·2 pI unit. Crystals suitable for X-ray diffraction analysis were prepared from acidic solutions using precipitation with ammonium sulfate. The hexagonal crystals diffracted X-rays to beyond 3·1 Å resolution and belonged to space group P61 (or P65) with cell constants a = b = 125·9 A ̊, c = 104·1 A ̊, γ = 120·0 °. The crystals likely contain two molecules in the asymmetric unit, giving a solvent content of 56% ( v w). Biochemical analysis of crystals indicated that both isoforms were present in approximately equimolar proportions. Full structure determination of the enzyme is underway. © 1992.

引用

页码：265 / 270

页数：6

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[1] REVISED 2.3-A STRUCTURE OF PORCINE PEPSIN - EVIDENCE FOR A FLEXIBLE SUBDOMAIN [J].