| Alternative title | Structural and catalytic properties of a deletion derivative (delta 133-157) of Escherichia coli adenylate kinase |
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| Download | - View final version: Structural and catalytic properties of a deletion derivative (Δ ₁₃₃₋₁₅₇) of Escherichia coli adenylate kinase (PDF, 2.5 MiB)
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| DOI | Resolve DOI: https://doi.org/10.1016/S0021-9258(18)99086-1 |
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| Author | Search for: Rose, Thierry; Search for: Brune, Martin; Search for: Wittinghofer, Alfred; Search for: Le Blay, Karine; Search for: Surewicz, Witold K.1; Search for: Mantsch, Henry H.1; Search for: Bârzu, Octavian; Search for: Gilles, Anne-Marie |
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| Affiliation | - National Research Council of Canada. NRC Steacie Institute for Molecular Sciences
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| Format | Text, Article |
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| Abstract | Escherichia coli adenylate kinase (AKe) as well as the enzyme from yeast and mitochondria differs from the muscle cytosolic variant (AK1) by an insertion of 25 amino acid residues that are missing in AK1. The extra sequence, highly homologous in "large" size variants, is situated between residues 133 and 157 in AKe. Removal of 25 codons in the corresponding adk gene resulted in expression of a modified form of adenylate kinase (Δ ₁₃₃₋₁₅₇ AKe) which still conserved 7% of the maximal activity of the wild-type protein. The apparent Kₘ for nucleotide substrates was increased by a factor of 4.6 (ADP), 23 (ATP) or 43 (AMP) in Δ ₁₃₃₋₁₅₇ AKe when compared with the wild-type enzyme. The secondary structure of Δ ₁₃₃₋₁₅₇ AKe, as well as its thermal stability were very similar to the parent protein. However, the deleted protein was much more sensitive than the wild-type enzyme to inactivation by trypsin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of trypsin digested Δ ₁₃₃₋₁₅₇ AKe revealed accumulation of several well defined fragments which were not observed in the case of wild-type enzyme. We conclude that the additional sequence, although necessary for expression of full activity in AKe, is not critical for catalysis. It is perhaps responsible for interaction of enzyme with other cellular components although a different mechanism of water shielding for large and small size variants of AK can be also envisaged. |
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| Publication date | 1991-06-15 |
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| Publisher | The American Society for Biochemistry and Molecular Biology, Inc.
Elsevier |
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| Licence | |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NPARC number | 23001319 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 6d318005-e122-4c01-b571-f41c639be9ed |
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| Record created | 2017-01-17 |
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| Record modified | 2021-04-30 |
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