Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase.
Protein ageing is often mediated by the formation of succinimide intermediates. These short-lived intermediates derive from asparaginyl deamidation and aspartyl dehydration and are rapidly converted into β-aspartyl or D-aspartyl residues. Here we report the presence of a highly stable succinimide intermediate in the glutaminase subunit of GMP synthetase from the hyperthermophile Methanocaldoccocus jannaschii. By comparing the biophysical properties of the wild-type protein and of several mutants, we show that the presence of succinimide increases the structural stability of the glutaminase subunit. The protein bearing this modification in fact remains folded at 100 °C and in 8 M guanidinium chloride. Mutation of the residue following the reactive asparagine provides insight into the factors that contribute to the hydrolytic stability of the succinimide. Our findings suggest that sequences that stabilize succinimides from hydrolysis may be evolutionarily selected to confer extreme thermal stability.
Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur, Bangalore 560064, India.
Sanjeev
S
Kumar
Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560012, India.
Sunita
S
Prakash
Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560012, India.
Kallol
K
Gupta
Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur, Bangalore 560064, India.
Aparna
A
Dongre
Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560012, India.
Padmanabhan
P
Balaram
Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur, Bangalore 560064, India.
Hemalatha
H
Balaram
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