CHARACTERIZATION OF RAT MAST-CELL GRANULE PROTEINS
CHARACTERIZATION OF RAT MAST-CELL GRANULE PROTEINS
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DOI：
10.1016/0003-9861(76)90292-7
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发表时间：
1976-01-01
影响因子：
3.9
通讯作者：
PRITZL, P
中科院分区：
文献类型：
--
作者：
LAGUNOFF, D;PRITZL, P
Mast cell granules free of membranes were isolated by differential centrifugation of water-lysed cells. The granules were extracted sequentially with 0.5, 1.0, and 2.0 M KCl. The 0.5 M fraction contained 95% of the N-acetyl-.beta.-glucosaminidase activity; this enzyme probably accounts for no more than 1% of the total granule protein. The 1.0 M fraction contained more than 80% of the granule chymotrypsin-like activity; the chymotrypsin-like enzyme was calculated to represent at least 15% of total granule protein. Heparin was found largely in the 1.0 M extract and in the residue after 2.0 M extraction. The heparin in both fractions had a molecular weight by gel exclusion chromatography considerably in excess of commercial porcine heparin. Acrylamide-gel electrophoresis of granules dissolved in 1% sodium dodecyl sulfate and reduced with dithiothreitol demonstrated 4 major protein bands. The 1.0 M fraction contained the most prominent, rapidly migrating band. The more slowly migrating, higher molecular weight bands appeared in grearer proportion in the 2.0 M and residue fractions. Autodigestion of the 1.0 M extract permitted purification of the mast cell chymotrypsin-like enzyme to specific activities as high as that of crystallized bovine pancreatic .alpha.-chymotrypsin. The mast cell chymotrypsin-like enzyme purified in this way migrated on dodecyl sulfate-gel electrophoresis as a single major band with an estimated molecular weight of 29,000.
