Preliminary-X-Ray Analysis of a Truncated Form of Recombinant Fructose-2,6-Bisphosphatase

Lee, Y. H., Lin, K., Okar, D., Alfano, N. L., Sarma, R., Pflugrath, J. W., Pilkis, S. J. (1994) Preliminary-X-Ray Analysis of a Truncated Form of Recombinant Fructose-2,6-Bisphosphatase. Journal of Molecular Biology, 235 (3). pp. 1147-1151. ISSN 0022-2836

URL: http://www.ncbi.nlm.nih.gov/pubmed/8289315

Abstract

The bisphosphatase domain of rat liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase and a C-terminal 30 amino acid truncated form were expressed in high yield in Escherichia coli and purified to homogeneity. The separately expressed bisphosphatase domain and its C-terminal truncated form had kinetic properties similar to the bisphosphatase of the intact bifunctional enzyme, but their turnover numbers were fourfold higher. The truncated enzyme crystallized in space group P1 with two molecules per asymmetric unit. The determined cell dimensions are: a = 41·9 Å, b = 43·5 Å, c = 57·6 Å, α = 95·2°, β = 99·3°, and γ = 106·2°. These crystals diffract to 2·0 Å resolution when exposed to synchrotron radiation and are suitable for crystallographic structure analysis.

Item Type: Paper
Uncontrolled Keywords: FRUCTOSE-2,6-BISPHOSPHATASE CLONING EXPRESSION PURIFICATION CRYSTALLIZATION HEPATIC 6-PHOSPHOFRUCTO-2-KINASE FRUCTOSE-2,6-BISPHOSPHATASE PHOSPHO GROUP DOMAIN MUTAGENESIS EXPRESSION BINDING
Subjects: Investigative techniques and equipment > cloning
Investigative techniques and equipment > assays > cloning

biotechnology > chromatography > protein purification
Investigative techniques and equipment > x ray crystallography
CSHL Authors:
Communities: CSHL labs
Depositing User: Matt Covey
Date Deposited: 04 May 2015 15:07
Last Modified: 04 May 2015 15:07
Related URLs:
URI: http://repository.cshl.edu/id/eprint/31478

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