EXPRESSION OF TRANSGLUTAMINASE-1 IN HUMAN EPIDERMIS
EXPRESSION OF TRANSGLUTAMINASE-1 IN HUMAN EPIDERMIS
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DOI：
10.1111/1523-1747.ep12612769
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发表时间：
1995-02-01
影响因子：
6.5
通讯作者：
STEINERT, PM
中科院分区：
文献类型：
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作者：
KIM, SY;CHUNG, SI;STEINERT, PM
为探讨膜相关或I型转氨酶（TGase 1）在人表皮中的表达及功能，我们制备了一种新的山羊抗人TGase 1抗体，该抗体针对纯化的具有活性的细菌表达的重组蛋白，通过免疫印迹和免疫沉淀方法，该抗体仅与表皮和培养的角质形成细胞中的TGase 1活性发生高度特异性反应，通过间接免疫荧光，抗体修饰了整个表皮，包括基底层，颗粒层具有一定的增强作用，然而，这些染色性质与广泛使用的市售TGase 1单克隆抗体的染色性质完全不同（称为B.C1），其装饰表皮的颗粒层，通过Western印迹，其仅微弱地鉴定TGase 1蛋白条带，但强烈识别一组15-20 kDa的条带，通过氨基酸分析和氨基酸测序，其中两个是小的富含脯氨酸（SPR）1和SPR 2蛋白，也在表皮和上皮组织中表达，连同用TGase 1蛋白和合成肽的一系列阻断实验，这些数据表明B.C1抗体的主要表位最可能位于这两种SPR蛋白的氨基末端而不是TGase 1上。现在需要进一步的研究来确定TGase 1在表皮发育和分化的不同阶段中的作用。
To explore the expression and function of the membrane-associated or type I transglutaminase (TGase1) in human epidermis, we have made a new antihuman TGase1 antibody in goats elicited against a purified active recombinant protein expressed in bacteria, By use of Western blotting and immunoprecipitation methods, the antibody reacted with high specificity with only the TGase1 activity of the epidermis and in cultured keratinocytes, By indirect immunofluorescence, the antibody decorated the entire epidermis, including the basal layer, with some potentiation of the granular layer, However, these staining properties are quite different from those of a widely used, commercially available TGase1 monoclonal antibody (termed B.C1), which decorates the granular layers of the epidermis, By Western blotting, it identifies the TGase1 protein band only weakly, but recognizes strongly a group of bands of 15-20 kDa, two of which by amino acid analysis and amino acid sequencing are the small proline-rich (SPR) 1 and SPR2 proteins, also expressed in epidermal and epithelial tissues, Together with a series of blocking experiments with TGase1 proteins and synthetic peptides, these data reveal that the major epitope of the B.C1 antibody most likely resides on the amino-terminus of these two SPR proteins rather than on TGase1. Further studies will now be necessary to determine the role(s) of TGase1 during the different stages of development and differentiation in the epidermis.