Evidence of eosinophil granule major basic protein in human placenta.
Evidence of eosinophil granule major basic protein in human placenta.
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DOI：
10.1084/jem.170.6.2051
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发表时间：
1989-12-01
期刊：
影响因子：
--
通讯作者：
Gleich GJ
中科院分区：
文献类型：
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作者：
Wasmoen TL;McKean DJ;Benirschke K;Coulam CB;Gleich GJ
一种与嗜酸性粒细胞主要碱性蛋白(GMBP)免疫化学相关的蛋白在孕妇血浆中的浓度升高，并通过免疫荧光定位于胎盘滋养层细胞。妊娠MBP(PMBP)与多克隆抗血清和一组14种小鼠单抗的反应性与gMBP无明显区别。本文报道了从人胎盘中分离纯化pMBP的方法：(A)SepharosemAb亲和层析，(B)6M盐酸胍缓冲液凝胶过滤，(C)反相高效液相色谱分离。纯化的pMBP和gMBP在生物化学上是不可区分的，因为它们都：(A)与DNA结合，(B)通过二硫键聚合并结合到载体蛋白上，(C)具有14,000的分子量，(D)具有大于10.6的等电点，(E)在二维凝胶中具有共价物，(F)在C18柱上进行反相高效液相色谱时的腔液，(G)经过三次不同的消化后具有相同的肽图，以及(H)具有部分氨基酸序列一致性。这种物理化学特性对于pMBP在人类胎盘形成中的作用具有重要的意义。
A protein immunochemically related to the eosinophil granule major basic protein (gMBP) is found in increased concentration in the plasma of pregnant women and has been localized to placental trophoblasts by immunofluorescence. Pregnancy MBP (pMBP) is indistinguishable from gMBP in its reactivity with polyclonal antisera and a panel of 14 mouse mAbs. We report the purification of pMBP from human placenta by: (a) affinity chromatography over mAb immobilized on Sepharose, (b) gel filtration in 6 M guanidine.HCl buffer, and (c) reversed-phase HPLC. Purified pMBP and gMBP are biochemically indistinguishable in that both: (a) bind to DNA, (b) polymerize and bind to carrier proteins via disulfide linkages, (c) have a molecular weight of 14,000, (d) have isoelectric points greater than 10.6, (e) comigrate in two-dimensional gels, (f) coelute during reversed-phase HPLC on C18 columns, (g) have identical peptide maps after three different digestions, and (h) have partial amino acid sequence identity. This physicochemical identity has important implications as to the role of pMBP in human placentation.