Purification to homogeneity of latent and active 58-kilodalton forms of human neutrophil collagenase.
Purification to homogeneity of latent and active 58-kilodalton forms of human neutrophil collagenase.
复制标题
纯化潜伏型和活性型 58 道尔顿人类中性粒细胞胶原酶的均质性。
DOI：
10.1021/bi00499a007
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发表时间：
1990
期刊：
影响因子：
2.9
通讯作者：
VanWart,HE
中科院分区：
文献类型：
--
作者：
Mookhtiar,KA;VanWart,HE
佛罗里达州塔拉哈西，佛罗里达州立大学化学系和分子生物物理研究所，32306收到1990年6月6日；修订稿件收到1990年8月9日摘要：潜在的和活性的58 kDa的人中性粒细胞胶原酶(HNC)已经纯化到均一。在苯甲基磺酰氟存在和不存在的情况下提取黄褐色的大衣，以产生分别含有潜伏性和活性HNC的粗制起始制剂。制备这些提取物和所有后续层析步骤所用的缓冲液含有浓度为0.5M或更高的氯化钠，0.05%的Brij-35，浓度为5 mM或更高的CaCl2，以及(如果可行)50QM的ZnS04以稳定HNC。在0.5M的氯化钠溶液中，用pH 7.5的活性红120-琼脂糖柱吸附棕黄色大衣提取液中的胶原酶活性，当氯化钠浓度增加到1。
Department of Chemistry and Institute of Molecular Biophysics, Florida State University, Tallahassee, Florida 32306 Received June 6, 1990; Revised Manuscript Received August 9, 1990 abstract: Latent and active 58-kDa forms of human neutrophil collagenase (HNC) have been purified to homogeneity. Buffy coats were extracted in the presence and absence of phenylmethanesulfonyl fluoride to generate crude starting preparations that contained latent and active HNC, respectively. The buffers used in preparing these extracts and for all subsequent chromatographic steps contained NaCl at a con-centration of 0.5 M or greater, 0.05% Brij-35, concentrations of CaCl2 of 5 mM or greater, and (when feasible) 50 qM ZnS04 to stabilize the HNC. The collagenase activity in the buffy coat extracts was adsorbed to a Reactive Red 120-agarose column at pH 7.5 in 0.5 M NaCl and was eluted when the NaCl concentration was increased to 1