A novel metalloproteinase associated with brain myelin membranes. Isolation and characterization.
A novel metalloproteinase associated with brain myelin membranes. Isolation and characterization.
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一种与脑髓磷脂膜相关的新型金属蛋白酶。
DOI：
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发表时间：
1989
期刊：
影响因子：
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通讯作者：
P. Glynn
中科院分区：
文献类型：
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作者：
Andrew Chantry;Norman A. Gregson;P. Glynn
脑髓鞘膜制剂含有金属蛋白酶活性，可降解髓鞘碱性蛋白(MBP)。该活性与从髓鞘中溶解的小扁豆凝集素结合糖蛋白有关，并可在洗涤剂3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate(CHAPS)存在下检测到。经DEAE-Sephacel、CM-Sepharose和Superose 6柱层析分离得到金属蛋白酶，经凝胶过滤和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法测得表观相对分子质量(MR)约为58000。MER值不受还原剂存在的影响，但经内切糖苷酶F处理后，其mR值降至约52,000。纯化的酶能裂解MBP中的许多键，但不产生三氯乙酸溶解产物。在纯化的酶或髓鞘膜消化的MBP中，两个主要的多肽可能是MBP1-73和MBP74-170。该酶在pH 7~9范围内具有活性，可被邻菲咯啉和二硫苏糖醇抑制，但不能被磷酰胺或丝氨酸或半胱氨酸蛋白酶抑制剂所抑制。组蛋白，但不是偶氮酶，也是该酶的底物。从它的酶和分子特性来看，髓鞘衍生的金属蛋白酶似乎有别于以前所描述的酶。
Brain myelin membrane preparations contain a metalloproteinase activity which degrades myelin basic protein (MBP). The activity was associated with lentil lectin-binding glycoproteins solubilized from myelin and could be detected in the presence of the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate (CHAPS). The metalloproteinase represented about 5% of this glycoprotein fraction and was isolated from it by chromatography on DEAE-Sephacel, CM-Sepharose, and Superose 6. The proteinase had an apparent relative molecular weight (Mr) of approximately 58,000 both by gel filtration and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Mr value was unaffected by the presence of reducing agents but was diminished to about 52,000 by treating the proteinase with endoglycosidase F. The purified proteinase cleaved many bonds in MBP but did not generate trichloroacetic acid-soluble products. Two major polypeptides, putatively MBP1-73 and MBP74-170, were prominent in digests of MBP by either the purified enzyme or myelin membranes. The proteinase was active between pH 7 and 9 and was inhibited by phenanthroline and dithiothreitol but not phosphoramidon or inhibitors of serine or cysteine proteinases. Histones, but not azocasein, also served as substrates for the proteinase. From its enzymic and molecular characteristics the myelin-derived metalloproteinase appears distinct from previously described enzymes.