title: Investigation of adenylate kinase from Escherichia coli and its interaction with nucleotides by Fourier transform infrared spectroscopy.
abstract: The secondary structure of adenylate kinase (EC 2.7.4.3) from E. coli was investigated under various conditions using Fourier transform infrared spectroscopy. The overall band contour of the conformation-sensitive amide I mode indicates that in HEPES buffer (pH 7.4) the major structure of the protein is alpha-helical. A more detailed estimate obtained from decomposition of the amide I band into its constituent component bands gives 50% alpha-helix, 26% beta-structure, 15% turns and loops, and about 9% nonrepetitive unordered structures. Binding of nucleotide (e.g., ATP) to the donor site decreases the beta-content and shifts the amide I band to higher wavenumbers, whereas binding of nucleotide (e.g., AMP) to the acceptor site does not produce any change in conformation of the protein. These results agree with the protection by ATP and lack of protection by AMP when adenylate kinase is digested with trypsin. The effect of protein denaturing agents and conditions (temperature, high pH, sodium dodecyl sulfate) on changes in the protein conformation as revealed by the conformation-sensitive amide I bands is discussed.
authors.affiliation: Department of Biochemistry, University of the Basque Country, Bilbao, Spain.
authors.forename: J L
authors.initials: JL
authors.lastname: Arrondo
authors.forename: A M
authors.initials: AM
authors.lastname: Gilles
authors.forename: O
authors.initials: O
authors.lastname: Bárzu
authors.forename: S
authors.initials: S
authors.lastname: Fermandjian
authors.forename: P W
authors.initials: PW
authors.lastname: Yang
authors.forename: H H
authors.initials: HH
authors.lastname: Mantsch
mesh_terms: D000263:Adenylate Kinase / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D005583:Fourier Analysis; D009711:Nucleotides / Q000378:metabolism*; D010770:Phosphotransferases / Q000378:metabolism*; D011487:Protein Conformation; D011489:Protein Denaturation; D013055:Spectrophotometry, Infrared
chemical_list: D009711:Nucleotides; D010770:Phosphotransferases; D000263:Adenylate Kinase