Structural and catalytic properties of a deletion derivative (delta 133-157) of Escherichia coli adenylate kinase

Download
  1. Get@NRC: Structural and catalytic properties of a deletion derivative (delta 133-157) of Escherichia coli adenylate kinase (Opens in a new window)
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titleThe Journal of Biological Chemistry
ISSN1083-351X
Volume266
Issue17
Pages1078110786
AbstractEscherichia 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 (delta 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 delta 133-157 AKe when compared with the wild-type enzyme. The secondary structure of delta 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 delta 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.
Publication date
Publishere American Society for Biochemistry and Molecular Biology, Inc.
Linkhttp://www.jbc.org/content/266/17/10781.abstract
LanguageEnglish
AffiliationNational Research Council Canada
Peer reviewedYes
NPARC number23001319
Export citationExport as RIS
Report a correctionReport a correction
Record identifier6d318005-e122-4c01-b571-f41c639be9ed
Record created2017-01-17
Record modified2017-01-17
Bookmark and share
  • Share this page with Facebook (Opens in a new window)
  • Share this page with Twitter (Opens in a new window)
  • Share this page with Google+ (Opens in a new window)
  • Share this page with Delicious (Opens in a new window)