Microtube batch protein crystallization: applications to human immunodeficiency virus type 2 (HIV-2) protease and human renin.
For therapeutically relevant targets, the evaluation of enzymes in complex with their inhibitors by cocrystallization and high resolution structural analysis has become a vital component of structure-driven drug design and development. Two approaches, hanging drop vapor diffusion and a novel microtube batch method, were utilized in parallel to grow crystals of recombinant HIV-2 protease and recombinant human renin in complex with inhibitors. In the case of HIV-2 protease in complex with a reduced amide inhibitor, crystallization was achieved only by the microbatch method. In the case of human renin, the addition of precipitant was required for crystal growth. The microbatch method described here is a useful supplementary or alternative approach for screening parameters and generating crystals suitable for high resolution structural analysis.
Pav
S
S
Department of Biochemistry, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut 06877.
Lubbe
K
K
Dô
F
F
Lamarre
D
D
Pargellis
C
C
Tong
L
L
D016282:Aspartic Acid Endopeptidases / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D016333:HIV Protease; D015498:HIV-2 / Q000201:enzymology*; D006801:Humans; D012083:Renin / Q000737:chemistry*
D016282:Aspartic Acid Endopeptidases; D016333:HIV Protease; C065184:p16 protease, Human immunodeficiency virus 2; D012083:Renin