title: HIV-1 protease variants from 100-fold drug resistant clinical isolates: expression, purification, and crystallization.
abstract: 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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, 540 E. Canfield Avenue, Detroit, MI 48201, USA.
authors.forename: John F
authors.initials: JF
authors.lastname: Vickrey
authors.forename: Bradley C
authors.initials: BC
authors.lastname: Logsdon
authors.forename: Gheorghe
authors.initials: G
authors.lastname: Proteasa
authors.forename: Sarah
authors.initials: S
authors.lastname: Palmer
authors.forename: Mark A
authors.initials: MA
authors.lastname: Winters
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Merigan
authors.forename: Ladislau C
authors.initials: LC
authors.lastname: Kovari
mesh_terms: 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
chemical_list: D003062:Codon; D017320:HIV Protease Inhibitors; D011993:Recombinant Fusion Proteins; D007532:Isoleucine; D016333:HIV Protease