Secondary structure and signal assignments of human-immunodeficiency-virus-1 protease complexed to a novel, structure-based inhibitor.
We report comprehensive NMR studies in solution of the human-immunodeficiency-virus (HIV)-1 protease. Stable solutions of the protease were obtained by complexing the protein to a designed cyclic urea inhibitor DMP 323. A variety of triple-resonance experiments provided essentially complete 1H, 13C and 15N NMR signal assignments of the protease. These assignments, together with short-range NOE constraints, coupling constants and hydrogen-exchange data, yielded the secondary structure of the protease in solution. The results reported herein open the way to the determination of the high-resolution three-dimensional solution structures of protease/inhibitor complexes, as well as to studies of protease dynamics and solvent interactions.
Yamazaki
T
T
Bone Research Branch, National Institute of Dental Research, NIH, Bethesda, MD 20892.
Nicholson
L K
LK
Torchia
D A
DA
Stahl
S J
SJ
Kaufman
J D
JD
Wingfield
P T
PT
Domaille
P J
PJ
Campbell-Burk
S
S
D000595:Amino Acid Sequence; D001381:Azepines; D003001:Cloning, Molecular; D004926:Escherichia coli; D016333:HIV Protease / Q000737:chemistry* / Q000378:metabolism*; D017320:HIV Protease Inhibitors / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011485:Protein Binding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014508:Urea / Q000031:analogs & derivatives* / Q000378:metabolism
D001381:Azepines; D017320:HIV Protease Inhibitors; D011994:Recombinant Proteins; D014508:Urea; D016333:HIV Protease; C085816:DMP 323