Secondary Mutations M36I and A71V in the Human Immunodeficiency Virus Type 1 Protease Can Provide an Advantage for the Emergence of the Primary Mutation D30N

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• 作者：José ; C. Clemente ; Reena Hemrajani ; Lisa E. Blum ; Maureen M. Goodenow ; and Ben M. Dunn
• 刊名：Biochemistry
• 出版年：2003
• 出版时间：December 30, 2003
• 年：2003
• 卷：42
• 期：51
• 页码：15029 - 15035
• 全文大小：258K
• 年卷期：v.42,no.51(December 30, 2003)
• ISSN：1520-4995

文摘

Development of resistance mutations in enzymatic targets of human immunodeficiency virus1 (HIV-1) hampers the ability to provide adequate therapy. Of special interest is the effect mutationsoutside the active site of HIV-1 protease have on inhibitor binding and virus viability. We engineeredprotease mutants containing the active site mutation D30N alone and with the nonactive site polymorphismsM36I and/or A71V. We determined the K_i values for the inhibitors nelfinavir, ritonavir, indinavir, KNI272,and AG1776 as well as the catalytic efficiency of the mutants. Single and double mutation combinationsexhibited a decrease in catalytic efficiency, while the triple mutant displayed catalytic efficiency greaterthan that of the wild type. Variants containing M36I or A71V alone did not display a significant changein binding affinities to the inhibitors tested. The variant containing mutation D30N displayed a 2-6-foldincrease in K_i for all inhibitors tested, with nelfinavir showing the greatest increase. The double mutantscontaining a combination of mutations D30N, M36I, and A71V displayed -0.5-fold to +6-fold changesin the K_i of all inhibitors tested, with ritonavir and nelfinavir most affected. Only the triple mutant showeda significant increase (>10-fold) in K_i for inhibitor nelfinavir, ritonavir, or AG-1776 displaying 22-, 19-,or 15-fold increases, respectively. Our study shows that the M36I and A71V mutations provide a greaterlevel of inhibitor cross-resistance combined with active site mutation D30N. M36I and A71V, when presentas natural polymorphisms, could aid the virus in developing active site mutations to escape inhibitorbinding while maintaining catalytic efficiency.