Solution structure of the cAMP-dependent protein kinase catalytic subunit and its contraction upon binding the protein kinase inhibitor peptide.
Small-angle X-ray scattering and Fourier transform infrared (FTIR) spectroscopy experiments have been completed on the catalytic subunit of the cAMP-dependent protein kinase. Measurements were made both with and without the protein kinase inhibitor peptide, PKI alpha(5-22)amide. Binding of the peptide results in an overall contraction of the structure that is characterized by a decrease of 9% in radius of gyration and about 16% in the maximum linear dimension. Both the secondary structure content of the protein/peptide complex, as determined by FTIR, and the solution structure of this binary complex, as determined by X-ray scattering, agree well with the structural characteristics of this complex as elucidated by the crystal structure [Knighton, D.R., Zheng, J., Ten Eyck, L. F., Ashford, V.A., Xuong, N.H., Taylor, S.S., & Sowadsi, J. M. (1991a) Science 253, 407-414]. Further, the contraction of the structure observed by X-ray scattering upon inhibitor peptide binding is not accompanied by any detectable change in secondary structure content of the kinase. We have modeled the contraction of the kinase upon inhibitor peptide binding as a simple rotation of the large and small lobes seen in the crystal structure such that the cleft between them is closed. For a substrate these changes would then allow catalysis to ensue. The hinge for this movement occurs around a glycine that is one of the protein kinase family consensus amino acids.
Isotope and Nuclear Chemistry Division, Los Alamos National Laboratory, New Mexico 87545.
G A
GA
Olah
R D
RD
Mitchell
T R
TR
Sosnick
D A
DA
Walsh
J
J
Trewhella
D000595:Amino Acid Sequence; D002384:Catalysis; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D000242:Cyclic AMP / Q000494:pharmacology*; D005583:Fourier Analysis; D005998:Glycine; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010455:Peptides / Q000378:metabolism*; D010802:Phylogeny; D011494:Protein Kinases / Q000737:chemistry* / Q000378:metabolism; D012542:Scattering, Radiation; D012996:Solutions; D013055:Spectrophotometry, Infrared; D014965:X-Rays
D010455:Peptides; D012996:Solutions; C061479:protein kinase inhibitor peptide; D000242:Cyclic AMP; D011494:Protein Kinases; D005998:Glycine