应用
来自牛胰腺的羧肽酶 A 被用于研究牛羧肽酶 A 的可溶和可活化形式在 大肠杆菌中的表达。来自牛胰腺的羧肽酶 A 也被用于研究鸵鸟羧肽酶前体的分离和部分特性。
用 Sigma 公司生产的羧肽酶研究了绿僵菌羧肽酶 A (MeCPA) 的特异性。MeCPA 是经过基因工程改造的，便于从重组蛋白的 C-末端去除多聚组氨酸标签。在体外还用于脱酪氨酸 α-微管蛋白，以诱导与乙基-N-苯氨基甲酸酯 (EPC) 琼脂糖的高亲和力。
生化/生理作用
羧肽酶分离自牛胰腺腺，是一种金属酶，每摩尔蛋白质含有 1 g 锌原子。对肽类和蛋白质中羧基末端肽键的水解进行催化。主要特异于芳香族和疏水侧链，如苯丙氨酸、色氨酸或亮氨酸。这种酶也表现出酯酶活性。可被 β-苯基丙酸盐和吲哚乙酸盐抑制。
单位定义
25°C、pH 7.5 下 1 个单位每分钟水解 1.0 μmol 马尿酸-L-苯丙氨酸。
制备说明
用苯甲基磺酰氟处理以消除胰蛋白酶和糜蛋白酶活性。透析和重结晶：加入甲苯的水性混悬液。
分析说明
通过 E 1%/278 测定蛋白
Bodo Wiesler et al.
The Plant journal : for cell and molecular biology, 32(6), 1023-1032 (2002-12-21)
Auxin controls the orientation of cortical microtubules in maize coleoptile segments. We used tyrosinylated alpha-tubulin as a marker to assess auxin-dependent changes in microtubule turnover. Auxin-induced tyrosinylated alpha-tubulin, correlated with an elevated sensitivity of growth to antimicrotubular compounds such as
Heiko Witt et al.
Nature genetics, 45(10), 1216-1220 (2013-08-21)
Chronic pancreatitis is an inflammatory disorder of the pancreas. We analyzed CPA1, encoding carboxypeptidase A1, in subjects with nonalcoholic chronic pancreatitis (cases) and controls in a German discovery set and three replication sets. Functionally impaired variants were present in 29/944
Kaia Kukk et al.
Journal of biotechnology, 231, 224-231 (2016-06-19)
Vertebrate prostaglandin H synthases (PGHSs) are membrane-bound disulphide-containing hemoglycoproteins. Therefore, eukaryotic expression systems are required for the production of recombinant PGHSs. Recently we announced the expression of human PGHS-2 (hPGHS-2) in the yeast Pichia pastoris. Here we report improved production
Matthew A Cottee et al.
Nature communications, 12(1), 5590-5590 (2021-09-24)
Excessive replication of Saccharomyces cerevisiae Ty1 retrotransposons is regulated by Copy Number Control, a process requiring the p22/p18 protein produced from a sub-genomic transcript initiated within Ty1 GAG. In retrotransposition, Gag performs the capsid functions required for replication and re-integration.
Facile discovery of surrogate cytokine agonists.
Yen, et al.
Cell, 185, 1414-1430 (2023)
实验方案
在测定羧肽酶A活性时，使用马尿酸-L-苯丙氨酸在254nm处进行连续分光光度法测定。羧肽酶A水解C末端残基上的肽键。
Carboxypeptidase A activity measured via continuous spectrophotometric rate determination assay with hippuryl-L-phenylalanine substrate.
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