Hawley-Nelson P, Berchtold M W, Huitfeldt H, Spiegel J, Yuspa S H
J Invest Dermatol. 1986 Feb;86(2):157-62. doi: 10.1111/1523-1747.ep12284197.
Skin calcium-binding protein (SCaBP) is a calcium binding protein purified from whole rat skin. It has a molecular weight of approximately 12,000 daltons but migrates at Mr 13,000 on sodium dodecyl sulfate (SDS)-polyacrylamide gels. On nitrocellulose blots of SDS-polyacrylamide gels, 6 different antisera to SCaBP reacted equally well with SCaBP and parvalbumin (PV), an 11,500-dalton calcium-binding protein purified from rat skeletal muscle, which also migrates at Mr 13,000 on SDS-polyacrylamide gels. Rabbit antiserum to muscle PV also recognized both PV and SCaBP, and either protein absorbed specific antibodies against either antigen from both types of antisera. Soluble protein extracts from whole adult rat and mouse skin contained a Mr 13,000 protein which was recognized on nitrocellulose blots of SDS gels by both antisera. Blots of extracts from epidermis, dermis, whole skin, and skin scraped on the dermal side to remove hypodermal tissue revealed that the Mr 13,000 PV/SCaBP cross-reacting antigen was restricted to the hypodermal tissue removed by scraping. Immunofluorescent staining of Bouin-fixed skin sections with these antisera confirmed the localization of PV/SCaBP to the panniculus carnosus, a hypodermal muscle layer. Newborn mouse skin does not contain this antigen. Additional polypeptides of Mr 10,500 and 12,000 on SDS gels of extracts from the epidermis of newborn and adult rats and mice were found to be immunoreactive with anti-SCaBP serum. These polypeptides were not recognized by the PV antiserum, and the reactivity of anti-SCaBP for these antigens was not absorbed by purified PV or SCaBP. Our results indicate that SCaBP is antigenically indistinguishable from PV and is localized in the adult rodent panniculus carnosus, and that antisera to SCaBP are polyspecific, recognizing epidermal proteins in addition to SCaBP/PV.
皮肤钙结合蛋白（SCaBP）是一种从大鼠全皮中纯化得到的钙结合蛋白。其分子量约为12,000道尔顿，但在十二烷基硫酸钠（SDS）-聚丙烯酰胺凝胶上的迁移率为Mr 13,000。在SDS-聚丙烯酰胺凝胶的硝酸纤维素印迹上，6种不同的针对SCaBP的抗血清与SCaBP和小白蛋白（PV）反应良好，PV是一种从大鼠骨骼肌中纯化得到的11,500道尔顿的钙结合蛋白，在SDS-聚丙烯酰胺凝胶上的迁移率也为Mr 13,000。针对肌肉PV的兔抗血清也识别PV和SCaBP，并且两种蛋白质都能从两种抗血清中吸收针对任何一种抗原的特异性抗体。成年大鼠和小鼠全皮的可溶性蛋白提取物中含有一种Mr 13,000的蛋白质，在SDS凝胶的硝酸纤维素印迹上能被两种抗血清识别。对表皮、真皮、全皮以及从真皮侧刮取以去除皮下组织的皮肤提取物进行印迹分析，结果显示Mr 13,000的PV/SCaBP交叉反应抗原仅限于刮取去除的皮下组织。用这些抗血清对Bouin固定的皮肤切片进行免疫荧光染色，证实了PV/SCaBP定位于肉膜，即一层皮下肌肉层。新生小鼠皮肤不含这种抗原。在新生大鼠和小鼠以及成年大鼠和小鼠的表皮提取物的SDS凝胶上，发现Mr 10,500和12,000的其他多肽与抗SCaBP血清有免疫反应性。这些多肽不被PV抗血清识别，并且抗SCaBP对这些抗原的反应性不会被纯化的PV或SCaBP吸收。我们的结果表明，SCaBP在抗原性上与PV无法区分，且定位于成年啮齿动物的肉膜，并且针对SCaBP的抗血清具有多特异性，除了SCaBP/PV外还能识别表皮蛋白。
