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

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Effect of mutation of lysine-128 of the large subunit of ribulose bisphosphate carboxylase/oxygenase from Anacystis nidulans. / Bainbridge, G.; Anralojc, P. J.; Madgwick, P. J. et al.
In: Biochemical Journal, Vol. 336, No. 2, 01.12.1998, p. 387-393.

Research output: Contribution to Journal/Magazine › Journal article › peer-review

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Bainbridge, G. ; Anralojc, P. J. ; Madgwick, P. J. et al. / Effect of mutation of lysine-128 of the large subunit of ribulose bisphosphate carboxylase/oxygenase from Anacystis nidulans. In: Biochemical Journal. 1998 ; Vol. 336, No. 2. pp. 387-393.

Bibtex

@article{f3a6d307ac0742469a8ea9ec8687fc6f,

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

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.",

author = "G. Bainbridge and Anralojc, {P. J.} and Madgwick, {P. J.} and Pitts, {J. E.} and Parry, {M. A J}",

year = "1998",

month = dec,

day = "1",

doi = "10.1042/bj3360387",

language = "English",

volume = "336",

pages = "387--393",

journal = "Biochemical Journal",

issn = "0264-6021",

publisher = "Portland Press Ltd.",

number = "2",

}

RIS

TY - JOUR

T1 - Effect of mutation of lysine-128 of the large subunit of ribulose bisphosphate carboxylase/oxygenase from Anacystis nidulans

AU - Bainbridge, G.

AU - Anralojc, P. J.

AU - Madgwick, P. J.

AU - Pitts, J. E.

AU - Parry, M. A J

PY - 1998/12/1

Y1 - 1998/12/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0032374056&partnerID=8YFLogxK

U2 - 10.1042/bj3360387

DO - 10.1042/bj3360387

M3 - Journal article

C2 - 9820816

AN - SCOPUS:0032374056

VL - 336

SP - 387

EP - 393

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -

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