Inactivation of pyroglutamyl aminopeptidase by N alpha-carbobenzoxy-L-pyroglutamyl chloromethyl ketone.
Inactivation of pyroglutamyl aminopeptidase by N alpha-carbobenzoxy-L-pyroglutamyl chloromethyl ketone.
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N α-苯甲氧酯-L-焦谷氨酰氯甲基酮灭活焦谷氨酰氨肽酶。
DOI：
10.1093/oxfordjournals.jbchem.a133490
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发表时间：
1981
影响因子：
2.7
通讯作者：
D. Tsuru
中科院分区：
文献类型：
--
作者：
K. Fujiwara;E. Matsumoto;T. Kitagawa;D. Tsuru
解淀粉芽孢杆菌的焦谷氨酰氨基肽酶[吡咯烷酮-羧酸肽酶：EC 3.4.11.8]被N α -碳苯氧基- l -焦谷氨酰氯甲基酮（Z-PGCK）快速且不可逆地失活。失活的二级速率常数为1.1 × 10(5) M-1。s-1，该值与clostripain-TLCK反应的值相当。在相同条件下，该氯甲基酮衍生物的d -异构体对酶几乎是惰性的。失活反应被一种劣质底物，焦戊酰缬氨酸的存在所阻止。pcmb失活酶被2-巯基乙醇可逆活化后，不能与Z-PGCK反应。这些结果表明，这种氯甲基酮衍生物作为亲和标记，可能与酶的活性位点半胱氨酸残基反应，正如报道的l -焦氨酰氯甲基酮。
Pyroglutamyl aminopeptidase [pyrrolidone-carboxylate peptidase: EC 3.4.11.8] from Bacillus amyloliquefaciens was inactivated rapidly and irreversibly by N alpha-carbobenzoxy-L-pyroglutamyl chloromethyl ketone (Z-PGCK). The second-order rate constant of the inactivation was 1.1 x 10(5) M-1.s-1, a value which is comparable to that of the clostripain-TLCK reaction. The D-isomer of this chloromethyl ketone derivative was almost inert toward the enzyme under the same conditions. The inactivation reaction was prevented by the presence of a poor substrate, pyroglutamyl-valine. The PCMB-inactivated enzyme, that was reversibly reactivated by 2-mercaptoethanol, failed to react with Z-PGCK. These results suggest that this chloromethyl ketone derivative reacts as an affinity label, presumably with the active site cysteinyl residue of the enzyme, as was reported for L-pyroglutamyl chloromethyl ketone.