Purification and properties of rat uterine procollagenase.
Purification and properties of rat uterine procollagenase.
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大鼠子宫原胶原酶的纯化和性质。
DOI：
10.1016/0003-9861(83)90032-2
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发表时间：
1983
影响因子：
3.9
通讯作者：
Jeffrey,JJ
中科院分区：
文献类型：
--
作者：
Roswit,WT;Halme,J;Jeffrey,JJ
从产后大鼠子宫细胞的单层培养物中纯化了胶原原酶。纯化中的关键步骤是将原胶原酶从含胎牛血清的培养基中结合到肝素-琼脂糖凝胶上，然后用极低浓度（5-10 nm）的硫酸葡聚糖洗脱。所得洗脱液含有8-10%的胶原原酶。通过在DEAE-Sepharose上的离子交换色谱、在AcA-44上的凝胶过滤和在blue-Sepharose上的色谱完成纯化。大鼠子宫前胶原酶经两种聚丙烯酰胺凝胶电泳系统、AcA-44层析和平衡沉降超电泳分析显示为Mr ~ 58，000的蛋白质双联体。酶原形式被胰蛋白酶转化为Mr ~ 48，000的活性酶双联体。在纯化过程中经常产生少量的活性酶，它们与胰蛋白酶活化的胶原酶在消化学上无法区分。用DEAE-Sepharose层析可将活性胶原酶与酶原分离。两种形式的酶原双联体可以通过AcA-44上的凝胶过滤部分分离，初步分析表明每种形式具有相等的胶原溶解活性。大鼠子宫胶原酶的氨基酸分析表明，它是显着不同的其他两种脊椎动物的胶原酶的组成是已知的。子宫酶原异常富含甘氨酸和羟基氨基酸，并且比人皮肤成纤维细胞胶原酶酸性大得多，这与两种分子的不同离子交换行为一致。使用天然重构豚鼠皮肤I型胶原原纤维作为底物，大鼠子宫胶原酶在37 °C下的比活性约为3000 μg胶原/min/mg。这种酶能切割变性的胶原蛋白，但不能攻击各种非胶原蛋白。
A procollagenase from monolayer cultures of postpartum rat uterine cells has been purified. The crucial step in the purification is the binding of the procollagenase from crude, fetal bovine serum-containing culture medium to heparin-Sepharose, followed by elution with extremely low concentrations (5–10 nm) of dextran sulfate. Resultant eluates contain 8–10% procollagenase. Purification is completed by ion-exchange chromatography on DEAE-Sepharose, gel filtration on AcA-44, and chromatography on blue-Sepharose. Rat uterine procollagenase appears as a protein doublet ofMr~ 58,000, as indicated by two polyacrylamide gel electrophoresis systems, by AcA-44 chromatography, and by equilibrium sedimentation ultracentrifugal analysis. The proenzyme forms are converted by trypsin to an active enzyme doublet ofMr~ 48,000. Small amounts of active enzyme, which are often generated during the purification, are electrophoretically indistinguishable from trypsin-activated collagenase. Active collagenase can be separated from the zymogen forms by DEAE-Sepharose chromatography. The two forms of the proenzyme doublet can be partially separated by gel filtration on AcA-44 and preliminary analysis indicates each has equal collagenolytic activity. The amino acid analysis of rat uterine collagenase reveals it to be markedly different from two other vertebrate collagenases whose composition is known. The uterine proenzyme is unusually rich in glycine and in the hydroxy amino acids and is considerably more acidic than the human skin fibroblast collagenase, consistent with the different ionexchange behavior of the two molecules. The specific activity of rat uterine collagenase at 37 °C is approximately 3000 μg collagen/min/mg, using native reconstituted guinea pig skin type I collagen fibrils as substrate. The enzyme cleaves denatured collagen, but fails to attack a variety of noncollagen proteins.
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II 型胶原蛋白对蝌蚪胶原酶降解 I 型胶原蛋白的双重作用。
DOI：
--
发表时间：
1981
期刊：
Collagen and Related Research
影响因子：
--
作者：
H. Sunada;T. Hayashi;H. Hori;Y. Nagai
通讯作者：
Y. Nagai
影响因子：
2.9
作者：
Y. Nagai;C. Lapière;J. Gross
通讯作者：
J. Gross
大鼠子宫外植体培养物中胶原酶的潜在形式和活性形式：孕激素对转化的调节。
影响因子：
3.9
作者：
Bernadette Tyree;Jouko Halme;John J. Jeffrey
通讯作者：
John J. Jeffrey
影响因子：
2.9
作者：
J. Jeffrey;J. Gross
通讯作者：
J. Gross
人皮肤成纤维细胞前胶原酶：有机汞和胰蛋白酶的激活机制。
影响因子：
2.9
作者：
Stricklin,GP;Jeffrey,JJ;Roswit,WT;Eisen,AZ
通讯作者：
Eisen,AZ