Expression, Purification, Crystallization, and Biochemical-Characterization of a Recombinant Protein Phosphatase

Zhuo, S., Clemens, J. C., Hakes, D. J., Barford, D., Dixon, J. E. (August 1993) Expression, Purification, Crystallization, and Biochemical-Characterization of a Recombinant Protein Phosphatase. J Biol Chem, 268 (24). pp. 17754-17761. ISSN 0021-9258

URL: http://www.ncbi.nlm.nih.gov/pubmed/8394350

Abstract

A protein phosphatase (PPase) from the bacteriophage lambda was overexpressed in Eschericha coli. The recombinant enzyme was purified to homogeneity yielding approximately 17 mg of enzyme from a single liter of bacterial culture. Biochemical characterization of the enzyme showed that it required Mn2+ or Ni2+ as an activator. The recombinant enzyme was active toward serine, threonine, and tyrosine phosphoproteins and phosphopeptides. Surprisingly, the bacterial histidyl phosphoprotein, NR(II), was also dephosphorylated by the lambda-PPase. The lambda-PPase shares a number of kinetic and structural properties with the eukaryotic Ser/Thr phosphatases, suggesting that the lambda-PPase will serve as a good model for structure-function studies. Crystallization of the recombinant purified lambda-PPase yielded monoclinic crystals. The crystals diffract to 4.0 angstrom when exposed to synchrotron x-ray radiation.

Item Type: Paper
Uncontrolled Keywords: CELLULAR-REGULATION CATALYTIC SUBUNIT PHOSPHOPROTEIN PHOSPHATASE TYROSINE-PHOSPHATASE BACTERIAL CHEMOTAXIS SKELETAL-MUSCLE RAT-LIVER PHOSPHORYLATION TYPE-1 KINASE
Subjects: organism description > bacteria > escherichia coli
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > protein phosphatase
CSHL Authors:
Communities: CSHL labs > Stillman lab
Depositing User: Matt Covey
Date: 25 August 1993
Date Deposited: 18 Apr 2016 20:00
Last Modified: 18 Apr 2016 20:00
Related URLs:
URI: http://repository.cshl.edu/id/eprint/32530

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