KINETIC CHARACTERIZATION AND CROSS-RESISTANCE PATTERNS OF HIV-1 PROTEASE MUTANTS SELECTED UNDER DRUG PRESSURE
KINETIC CHARACTERIZATION AND CROSS-RESISTANCE PATTERNS OF HIV-1 PROTEASE MUTANTS SELECTED UNDER DRUG PRESSURE
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DOI：
10.1021/bi00029a002
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发表时间：
1995-07-25
期刊：
影响因子：
2.9
通讯作者：
ERICKSON, JW
中科院分区：
文献类型：
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作者：
GULNIK, SV;SUVOROV, LI;ERICKSON, JW
基于使用抑制剂A-77003的体外选择实验的结果，产生了11种不同的重组、耐药性HIV-1蛋白酶（HIV PR）突变体-R8 Q、V32 I、M46 I、V82 A、V82 F、V82 I、184 V、V321/I84 V、M46 I/V82 F、M46 I/I84 V和V321/K451/F53 L/A71 V/I84 V/L 89 M，A-84538和KNI-272。测定突变型和野生型（WT）酶的动力学参数沿着对抑制剂A-77003、A-84538、KNI-272、L-735，524和Ro 31 -8959的抑制常数（Ki）。突变体的催化效率k（cat）/K-m相对于WT降低1.2-14.8倍，主要是由于K-m升高。特定突变对Ki值的影响对于抑制剂和突变酶都是独特的。引入了一个新的性质，称为活力，定义为比率（K（i）k（cat）/K-m）（突变体）/（K（i）k（cat）/K-m）（WT），以比较不同突变体在给定抑制剂存在下的选择优势。体外选择研究期间出现的突变通常会观察到高活力值。动力学模型沿着的突变体面板这里描述的应该是有用的评估和预测模式的耐药HIV PR抑制剂，并可能有助于抑制剂组合的选择，以打击耐药性。
Eleven different recombinant, drug-resistant HIV-1 protease (HIV PR) mutants-R8Q, V32I, M46I, V82A, V82F, V82I, 184V, V321/I84V, M46I/V82F, M46I/I84V, and V321/K451/F53L/A71V/I84V/ L89M-were generated on the basis of results of in vitro selection experiments using the inhibitors A-77003, A-84538, and KNI-272. Kinetic parameters of mutant and wild-type (WT) enzymes were measured along with inhibition constants (K-i) toward the inhibitors A-77003, A-84538, KNI-272, L-735,524, and Ro31-8959. The catalytic efficiency, k(cat)/K-m, for the mutants decreased relative to WT by a factor of 1.2-14.8 and was mainly due to the elevation of K-m. The effects of specific mutations on K-i values were unique with respect to both inhibitor and mutant enzyme. A new property, termed vitality, defined as the ratio (K(i)k(cat)/K-m)(mutant)/(K(i)k(cat)/K-m)(WT) was introduced to compare the selective advantage of different mutants in the presence of a given inhibitor. High vitality values were generally observed with mutations that emerged during in vitro selection studies. The kinetic model along with the panel of mutants described here should be useful for evaluating and predicting patterns of resistance for HIV PR inhibitors and may aid in the selection of inhibitor combinations to combat drug resistance.