脉冲辐解技术在蛋白质研究中的应用：胰凝乳蛋白酶和胰蛋白酶。
Application of pulse radiolysis to the study of proteins: chymotrypsin and trypsin.
作者信息
Faraggi M, Klapper M H, Dorfman L M
出版信息
Biophys J. 1978 Oct;24(1):307-17. doi: 10.1016/S0006-3495(78)85379-X.
Abstract
The one-electron reduction of chymotrypsin, trypsin, and their zymogens have been studied by pulse radiolysis. The optical spectra of the transient products from the two active enzymes display a pH-dependent band at 360 nm, associated with the histidine-electron adduct. The yield of the histidyl radical as a function of pH is consistent with a pK(a) less than 4.5, which suggests that the radical is located at the enzyme active site. The histidines of the proenzymes chymotrypsinogen and trypsinogen are unreactive towards the hydrated electron. We conclude that formation of the histidine-electron adduct at the serine protease active site is sensitive to the physical alterations which accompany protease activation.
摘要
通过脉冲辐解研究了胰凝乳蛋白酶、胰蛋白酶及其酶原的单电子还原。两种活性酶瞬态产物的光谱在360nm处显示出一个pH依赖性谱带，与组氨酸-电子加合物相关。组氨酸自由基的产率作为pH的函数，其pK(a)小于4.5，这表明该自由基位于酶的活性位点。胰凝乳蛋白酶原和胰蛋白酶原的组氨酸对水合电子无反应。我们得出结论，丝氨酸蛋白酶活性位点处组氨酸-电子加合物的形成对蛋白酶激活时伴随的物理变化敏感。
相似文献
1
Application of pulse radiolysis to the study of proteins: chymotrypsin and trypsin.脉冲辐解技术在蛋白质研究中的应用：胰凝乳蛋白酶和胰蛋白酶。
Biophys J. 1978 Oct;24(1):307-17. doi: 10.1016/S0006-3495(78)85379-X.
2
Gamma-radiolysis of trypsin, chymotrypsin, and chymotrypsinogen when associated with DNA.胰蛋白酶、胰凝乳蛋白酶和胰凝乳蛋白酶原与DNA结合时的γ辐射分解
Radiat Res. 1974 Mar;57(3):395-402.
3
Zymogen activation in serine proteinases. Proton magnetic resonance pH titration studies of the two histidines of bovine chymotrypsinogen A and chymotrypsin Aalpha.丝氨酸蛋白酶中的酶原激活。牛胰凝乳蛋白酶原A和α-胰凝乳蛋白酶的两个组氨酸的质子磁共振pH滴定研究。
Biochemistry. 1978 Oct 31;17(22):4627-40. doi: 10.1021/bi00615a008.
4
Effect of ultrasound on activation of serine proteases precursors.超声对丝氨酸蛋白酶前体激活的影响。
Ultrason Sonochem. 2005 Feb;12(3):219-23. doi: 10.1016/j.ultsonch.2003.10.012.
5
Gamma-irradiated chymotrypsin-like proteins. I. Structural changes.γ射线辐照的类胰凝乳蛋白酶蛋白。I. 结构变化
Int J Radiat Biol Relat Stud Phys Chem Med. 1977 Feb;31(2):131-44. doi: 10.1080/09553007714550151.
6
The pulse radiolysis and inactivation of trypsin.胰蛋白酶的脉冲辐解与失活
Int J Radiat Biol Relat Stud Phys Chem Med. 1971;20(5):447-59. doi: 10.1080/09553007114551351.
7
Catalysis by serine proteases and their zymogens. A study of acyl intermediates by circular dichroism.丝氨酸蛋白酶及其酶原的催化作用。通过圆二色性对酰基中间体的研究。
Biochemistry. 1975 Nov 18;14(23):5088-94. doi: 10.1021/bi00694a010.
8
Properties of the His57-Asp102 dyad of rat trypsin D189S in the zymogen, activated enzyme, and alpha1-proteinase inhibitor complexed forms.大鼠胰蛋白酶D189S的His57-Asp102二元体在酶原、活化酶和α1-蛋白酶抑制剂复合形式下的特性。
Arch Biochem Biophys. 1999 Feb 15;362(2):254-64. doi: 10.1006/abbi.1998.1035.
9
Dynamic and structural properties of the calcium binding site of bovine serine proteases and their zymogens. A multinuclear nuclear magnetic resonance and stopped-flow study.牛丝氨酸蛋白酶及其酶原钙结合位点的动态和结构特性。一项多核磁共振和停流研究。
J Mol Biol. 1985 Sep 5;185(1):201-7. doi: 10.1016/0022-2836(85)90191-3.
10
Flexibility in the specificity site of serine proteases.丝氨酸蛋白酶特异性位点的灵活性。
Biochemistry. 1976 May 18;15(10):2212-9. doi: 10.1021/bi00655a029.
引用本文的文献
1
Aggregation of Albumins under Reductive Radical Stress.在还原自由基应激下白蛋白的聚集。
Int J Mol Sci. 2024 Aug 19;25(16):9009. doi: 10.3390/ijms25169009.
本文引用的文献
1
Rate constants of hydrated electron reactions with peptides and proteins.水合电子与肽和蛋白质反应的速率常数。
Radiat Res. 1967 May;31(1):8-26.
2
The study of alpha-chymotrypsin by x-ray diffraction. The Third CIBA Medal Lecture.通过X射线衍射对α-胰凝乳蛋白酶的研究。第三届西巴奖章讲座。
Biochem J. 1969 Apr;112(3):261-8. doi: 10.1042/bj1120261.
3
Structure of crystalline -chymotrypsin. II. A preliminary report including a hypothesis for the activation mechanism.结晶胰凝乳蛋白酶的结构。II. 初步报告，包括激活机制的假说。
J Mol Biol. 1968 Jul 14;35(1):143-64. doi: 10.1016/s0022-2836(68)80043-9.
4
Three-dimensional structure of tosyl-elastase.甲苯磺酰弹性蛋白酶的三维结构
Nature. 1970 Feb 28;225(5235):811-6. doi: 10.1038/225811a0.
5
The pulse radiolysis and inactivation of trypsin.胰蛋白酶的脉冲辐解与失活
Int J Radiat Biol Relat Stud Phys Chem Med. 1971;20(5):447-59. doi: 10.1080/09553007114551351.
6
Reactions of proteins with hydrated electrons: the effect of conformation on the reaction rate constant.蛋白质与水合电子的反应：构象对反应速率常数的影响。
Int J Radiat Biol Relat Stud Phys Chem Med. 1967;13(2):195-7. doi: 10.1080/09553006814550111.
7
Reductive deamination of oligopeptides by solvated electrons in aqueous solution.水溶液中溶剂化电子对寡肽的还原脱氨作用。
Radiat Res. 1971 Nov;48(2):244-55.
8
Structure of crystalline -chymotrypsin. V. The atomic structure of tosyl- -chymotrypsin at 2 A resolution.结晶胰凝乳蛋白酶的结构。V. 对甲苯磺酰基-胰凝乳蛋白酶在2埃分辨率下的原子结构
J Mol Biol. 1972 Jul 21;68(2):187-240. doi: 10.1016/0022-2836(72)90210-0.
9
Fast consecutive radical processes within the ribonuclease molecule in aqueous solution. II. Reaction with OH radicals and hydrated electrons.
Biochim Biophys Acta. 1972 Jul 13;276(1):124-42. doi: 10.1016/0005-2744(72)90014-9.
10
High resolution nuclear magnetic resonance studies of the active site of chymotrypsin. I. The hydrogen bonded protons of the "charge relay" system.胰凝乳蛋白酶活性位点的高分辨率核磁共振研究。I. “电荷中继”系统的氢键质子。
J Mol Biol. 1974 Jul 5;86(3):519-40. doi: 10.1016/0022-2836(74)90178-8.
Zotero 插件