MAST-CELL HETEROGENEITY - 2-DIMENSIONAL GEL-ELECTROPHORETIC ANALYSES OF RAT PERITONEAL AND INTESTINAL MUCOSAL MAST-CELLS
MAST-CELL HETEROGENEITY - 2-DIMENSIONAL GEL-ELECTROPHORETIC ANALYSES OF RAT PERITONEAL AND INTESTINAL MUCOSAL MAST-CELLS
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DOI：
10.1002/eji.1830200911
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发表时间：
1990-09-01
影响因子：
5.4
通讯作者：
BEFUS, AD
中科院分区：
文献类型：
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作者：
ABE, T;SWIETER, M;BEFUS, AD
从感染巴西尼波圆线虫的大鼠中纯化出腹膜肥大细胞（PMC）和肠粘膜肥大细胞（IMCC）。两种肥大细胞亚型的总体蛋白质成分通过二维凝胶电泳（IEF）在第一维和SDS-PAGE（10%）在第二维进行分析，然后进行银染色。PMC有7个优势碱性蛋白（PB2-pI 9-9.5），估计分子质量在26 ~ 37 kDa之间；还有80 ~ 90个中性或酸性蛋白，大多数pI 6 ~ 7.5，估计分子质量在20 ~ 100 kDa之间。所有的基本蛋白都是颗粒相关的。PB6 （29 kDa）、PB7 （28 kDa）和PB8 （RMCP I, 26 kDa）三个碱性蛋白结合[3H]二异丙基氟磷酸（DFP），表明它们是丝氨酸蛋白酶。然而，只有PB8与RCMP i抗体反应。另一种基本成分（< 14 kDa），可能是PB6、PB7或PB8的降解产物，也与[3H]DFP结合。相比之下，IMMC具有9个碱性蛋白（IB1-9），总体而言，它们比PMC酸性更强（pI约为8.5-9）。4种主要碱性蛋白（IB6-9）均为24 kDa，但在等电点上略有不同。这些和另一个46-kDa的基本成分（IB-2）与RCMP II抗体和结合[3H]DFP抗体反应。IMMC中未发现其他dfp结合蛋白。尽管PMC中的基本颗粒相关蛋白与IMMC中的基本蛋白之间存在显著差异，但IMMC中中性酸性范围内的斑点大部分相似，中性酸性范围内的斑点在两个肥大细胞亚群中大部分相似，尽管一些斑点的数量差异很明显。因此，来自腹腔和肠粘膜的大鼠肥大细胞群在其具有碱性pI的蛋白质成分中表现出明显的异质性，包括具有丝氨酸蛋白酶活性的颗粒相关蛋白。
Peritoneal mast cells (PMC) and intestinal mucosal mast cells (IMCC) were purified from rats infected with the nematode Nippostrongylus brasiliensis. Overall protein constituents of both mast cell subtypes were analyzed by two-dimensional gel electrophoresis using either (IEF) in the first dimension and SDS-PAGE (10%) in the second dimension followed by silver staining. PMC had seven dominant basic proteins (PB2-pI 9-9.5) with estimated molecular masses of 26 to 37 kDa, as well as 80 to 90 neutral or acidic proteins, most of which had pI 6 to 7.5 and estimated molecular masses of 20 to 100 kDa. All the basic proteins were granule-associated. Three basic proteins, PB6 (29 kDa), PB7 (28 kDa) and PB8 (RMCP I, 26 kDa), bound [3H]diisopropyl fluorophosphate (DFP), suggesting that they are serine proteases. However, only PB8 was reactive with antibodies to RCMP I. Another basic component (< 14 kDa), perhaps a degradation product of PB6, PB7 or PB8, also bound [3H]DFP. By comparison, IMMC possessed nine basic proteins (IB1-9) and, in general, they were more acidic (pI about 8.5-9) than those of PMC. Four major basic proteins (IB6-9) were all 24 kDa but were slightly different in isoelectric points. These and another 46-kDa basic component (IB-2) were reactive with antibodies to RCMP II and bound [3H]DFP. There were no other DFP-binding proteins in IMMC. In spite of remarkable differences between basic granule-associated proteins in PMC and basic proteins in IMMC, spots in the neutral-acidic range were for the most part similar proteins in IMMC, spots in the neutral-acidic range were for the most part similar in the two mast cell subsets, although quantitative differences were evident for some spots. Thus, rat mast cell populations from the peritoneal cavity and intestinal mucosa exhibit marked heterogeneity in their protein constituents with basic, pI, including in their granule-associated proteins with serine protease activity.