HIV-1 protease variants from 100-fold drug resistant clinical isolates: expression, purification, and crystallization.
High-resolution X-ray crystallographic structures of HIV-1 protease clinical variants complexed with licensed inhibitors are essential to understanding the fundamental cause of protease drug resistance. There is a need for structures of naturally evolved HIV-1 proteases from patients failing antiretroviral therapy. Here, we report the expression, purification, and crystallization of clinical isolates of HIV-1 protease that have been characterized to be more than 100 times less susceptible to US FDA approved protease inhibitors.
Vickrey
John F
JF
Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, 540 E. Canfield Avenue, Detroit, MI 48201, USA.
Logsdon
Bradley C
BC
Proteasa
Gheorghe
G
Palmer
Sarah
S
Winters
Mark A
MA
Merigan
Thomas C
TC
Kovari
Ladislau C
LC
D000595:Amino Acid Sequence; D003062:Codon / Q000235:genetics; D003460:Crystallization; D024882:Drug Resistance, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D016333:HIV Protease / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D017320:HIV Protease Inhibitors / Q000494:pharmacology; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D007532:Isoleucine / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D017510:Protein Folding; D020673:Protein Renaturation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D003062:Codon; D017320:HIV Protease Inhibitors; D011993:Recombinant Fusion Proteins; D007532:Isoleucine; D016333:HIV Protease