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Effect of mutation of lysine-128 of the large subunit of ribulose bisphosphate carboxylase/oxygenase from Anacystis nidulans

Bainbridge, G. and Anralojc, P. J. and Madgwick, P. J. and Pitts, J. E. and Parry, M. A J (1998) Effect of mutation of lysine-128 of the large subunit of ribulose bisphosphate carboxylase/oxygenase from Anacystis nidulans. Biochemical Journal, 336 (2). pp. 387-393. ISSN 0264-6021

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Abstract

The contribution of lysine-128 within the active site of Anacystis nidulans D-ribulose 1,5-bisphosphate carboxylase/oxygenase was investigated by the characterization of mutants in which lysine-128 was replaced with arginine, glycine, glutamine, histidine or aspartic acid. Mutated genes encoding the Rubisco large subunit were expressed in Escherichia coli and the resultant polypeptides assembled into active complexes. All of the mutant enzymes had a lower affinity for ribulose 1,5-bisphosphate (RuBP) and lower rates of carboxylation. Substitution of lysine-128 with glutamine, histidine or aspartic acid decreased the specificity factor and led to the production of an additional monophosphate reaction product. We show that this product results from the loss of the phosphate from C-1 of RuBP, most probably by β-elimination from the 2,3-enediolate derivative of RuBP. The results confirm that lysine-128 is important in determining the position of the essential ε-amino group of lysine-334 within the active site and in loop dynamics. This further demonstrates that residues remote from the active site can be manipulated to modify catalytic function.

Item Type: Article
Journal or Publication Title: Biochemical Journal
Subjects:
Departments: Faculty of Science and Technology > Lancaster Environment Centre
ID Code: 76033
Deposited By: ep_importer_pure
Deposited On: 21 Oct 2015 06:04
Refereed?: Yes
Published?: Published
Last Modified: 21 Sep 2017 01:15
Identification Number:
URI: http://eprints.lancs.ac.uk/id/eprint/76033

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