Cloning, purification, and crystallization of Escherichia coli cystathionine beta-lyase

被引：15

作者：

Laber, B

Clausen, T

Huber, R

Messerschmidt, A

Egner, U

MullerFahrnow, A

Pohlenz, HD

机构：

[1] MAX PLANCK INST BIOCHEM, D-82152 MARTINSRIED, GERMANY

[2] HOECHST SCHERING AGREVO GMBH, D-13476 BERLIN, GERMANY

[3] SCHERING AG, D-13342 BERLIN, GERMANY

来源：

FEBS LETTERS | 1996年 / 379卷 / 01期

关键词：

protein crystallization; X-ray crystallography; cystathionin beta-lyase; cystathionase; methionine biosynthesis; pyridoxal 5'-phosphate; Escherichia coli;

D O I：

10.1016/0014-5793(95)01499-3

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 metC gene coding for cystathionine beta-lyase of Escherichia coli has been cloned and used to construct an over-producing E, coli strain. An efficient purification scheme has been developed and the purified enzyme has been crystallized by the hanging drop vapour diffusion method using either ammonium sulfate or polyethyleneglycol 400 as precipitating agent. The crystals belong to the orthorombic space group C222(1). Their mit cell parameters are a = 60.9 Angstrom, b = 154.7 Angstrom and c = 152.7 Angstrom. Consideration of the possible values of V-M accounts for the presence of one dimer per asymmetric unit, The crystals are suitable for X-ray analysis and a complete native date set to 1.83 Angstrom resolution has been collected using synchrotron radiation.

引用

页码：94 / 96

页数：3

共 16 条

[1]

ALEXANDER FW, 1994, EUR J BIOCHEM, V219, P950

[2] 3-DIMENSIONAL STRUCTURE OF TYROSINE PHENOL-LYASE [J].

ANTSON, AA ;

DEMIDKINA, TV ;

GOLLNICK, P ;

DAUTER, Z ;

VONTERSCH, RL ;

LONG, J ;

BEREZHNOY, SN ;

PHILLIPS, RS ;

HARUTYUNYAN, EH ;

WILSON, KS .

BIOCHEMISTRY, 1993, 32 (16) :4195-4206

[3] EVOLUTION IN BIOSYNTHETIC PATHWAYS - 2 ENZYMES CATALYZING CONSECUTIVE STEPS IN METHIONINE BIOSYNTHESIS ORIGINATE FROM A COMMON ANCESTOR AND POSSESS A SIMILAR REGULATORY REGION [J].

BELFAIZA, J ;

PARSOT, C ;

MARTEL, A ;

DELATOUR, CB ;

MARGARITA, D ;

COHEN, GN ;

SAINTGIRONS, I .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (04) :867-871

[4]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[5]

JANSONIUS JN, 1987, BIOL MACROMOL, V3, P187

[6] PYRIDOXAL PHOSPHATE-DEPENDENT ENZYMES [J].

JOHN, RA .

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1995, 1248 (02) :81-96

[7] PYRIDOXAL 5'PHOSPHATE BINDING-SITE OF ESCHERICHIA-COLI BETA CYSTATHIONASE AND CYSTATHIONINE GAMMA-SYNTHASE COMPARISON OF THEIR SEQUENCES [J].

MARTEL, A ;

DELATOUR, CB ;

LEGOFFIC, F .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1987, 147 (02) :565-571

[8]

MATHEWS BW, 1968, J MOL BIOL, V33, P491

[9]

MILES EW, 1991, ADV ENZYMOL RAMB, V64, P93

[10]

PARSOT C, 1987, MICROBIOL SCI, V9, P258

← 1 2 →