Graduate Program of Biochemical Regulation, Graduate School of Bioagricultural Sciences, Nagoya University, Japan.
Cytotechnology. 2000 Jul;33(1-3):167-74. doi: 10.1023/A:1008186912975.
Laminins are a family of glycoproteins composed of alpha,beta and gamma chains. Five alpha(alpha1-alpha5), three beta (beta1-beta3) and twogamma (gamma1 and gamma2) chains have been cloned fromhuman and their replaceable assembly into heterotrimers producesthe variety of laminins. Reverse transcription-polymerase chainreaction of mRNAs showed that human keratinocytes express thealpha3, alpha5, beta1, beta3, gamma1 andgamma2 genes at high level among the ten cloned lamininchains. Western blot and immunoprecipitation of the cell lysatewith antiserum directed against mouse laminin-1(alpha1beta1gamma1) detected two trimers with thecomposition of alphaxbeta1gamma1 (probablylaminin-10 with the composition of alpha5beta1gamma1and alphaybeta1gamma1. Meanwhile, antiserum directedagainst a synthetic peptide of human alpha3 detected onlyalpha3beta3gamma2 trimer (laminin-5). We thus show thatkeratinocytes produce three heterotrimeric laminins. We couldnot detect the assembly of alpha3 with beta1 and gamma1chains to form alpha3beta1gamma1 (laminin-6) in keratinocytes.
层粘连蛋白是由α、β和γ链组成的糖蛋白家族。已从人源中克隆出五种α(α1-α5)、三种β(β1-β3)和两种γ(γ1 和γ2)链,它们的可替换组装成异三聚体产生多种层粘连蛋白。mRNA 的逆转录-聚合酶链反应显示,在十种已克隆的层粘连蛋白链中,人角质形成细胞高水平表达α3、α5、β1、β3、γ1 和γ2 基因。用针对小鼠层粘连蛋白-1(α1β1γ1)的抗血清对细胞裂解物进行 Western blot 和免疫沉淀,检测到两种αxβ1γ1 组成的三聚体(可能是α5β1γ1 和αyβ1γ1 组成的层粘连蛋白-10,同时,针对人α3 的合成肽的抗血清仅检测到α3β3γ2 三聚体(层粘连蛋白-5)。因此,我们表明角质形成细胞产生三种异三聚体层粘连蛋白。我们未能在角质形成细胞中检测到α3 与β1 和γ1 链组装形成α3β1γ1(层粘连蛋白-6)。