Cloning, purification, and crystallization of Escherichia coli cystathionine β‐lyase

FEBS Letters

Laber, Bernd²  Pohlenz, Hans-Dieter²  Clausen, Tim³  Egner, Ursula¹  Huber, Robert³  Messerschmidt, Albrecht³  Müller-Fahrnow, Anke¹ 

[1] Schering AG, Muellerstr. 170-178, D-13342 Berlin, Germany;Hoechst Schering AgrEvo GmbH, Gollanczstr. 71, D-13476 Berlin, Germany;Max-Planck-Institut für Biochemie, Am Klopferspitz 18a, D-82152 Martinsried, Germany

关键词: Protein crystallization;    X-Ray crystallography;    Cystathionin β-lyase;    Cystathionase;    Methionine biosynthesis;    Pyridoxal 5′-phosphate;    Escherichia coli;    CL;    cystathionin β-lyase;    FPLC;    fast performance liquid chromatography;    MES;    2-(N-morpholino)propanesulfonic acid;    PCR;    polymerase chain reaction;    PEG;    polyethylenglycol;    PLP;    pyridoxal 5′-phosphate;    V M;    Matthews-parameter (protein volume per mass unit in the crystal);   

DOI  :  10.1016/0014-5793(95)01499-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The metC gene coding for cystathionine β-lyase of Escherichia coli has been cloned and used to construct an overproducing 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 C222₁. Their unit cell parameters are . 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 Å resolution has been collected using synchrotron radiation.

【 授权许可】

Unknown