Lin B, Saito M, Sakakibara Y, Hayashi Y, Yanagisawa M, Iwamori M
Department of Biochemistry and Molecular Biology, Graduate School of Medicine, University of Tokyo, Japan.
Arch Biochem Biophys. 2001 Apr 15;388(2):207-15. doi: 10.1006/abbi.2001.2303.
We cloned three members of a GDP-fucose:beta-galactoside alpha1,2-fucosyltransferase (alpha1,2-fucosyltransferase) family, MFUT-I, -II, and -III, from a cDNA of murine small intestine, and determined their enzymatic properties after transfection of the genes into COS-7 cells, and their expression in murine tissues by Northern blotting. MFUT-I, -II, and -III exhibited sequence homology with the human H (78.4%), Se (79.0%), and Sec1 (74.9%) gene products, respectively. COS-7 cells transfected with MFUT-I and -II exhibited alpha1,2-fucosyltransferase activity and the best acceptor substrate for both gene products was GA1 to yield a fucosyl GA1 structure, but no activity was detected in COS-7 cells with MFUT-III. MFUT-II yielded a 3.5-kb mRNA transcript in several tissues, whereas MFUT-I and -III were predominantly expressed in epididymis and testis, respectively. The administration of microbes into germ-free mice resulted in a rapid increase of the MFUT-II gene (Se gene) for the synthesis of fucosyl GA1 in the intestine.
我们从小鼠小肠的cDNA中克隆了GDP-岩藻糖:β-半乳糖苷α1,2-岩藻糖基转移酶(α1,2-岩藻糖基转移酶)家族的三个成员,MFUT-I、-II和-III,并在将这些基因转染到COS-7细胞后测定了它们的酶学性质,通过Northern印迹法检测了它们在小鼠组织中的表达。MFUT-I、-II和-III分别与人H(78.4%)、Se(79.0%)和Sec1(74.9%)基因产物表现出序列同源性。用MFUT-I和-II转染的COS-7细胞表现出α1,2-岩藻糖基转移酶活性,这两种基因产物的最佳受体底物都是GA1,以产生岩藻糖基化的GA1结构,但在转染MFUT-III的COS-7细胞中未检测到活性。MFUT-II在多个组织中产生3.5 kb的mRNA转录本,而MFUT-I和-III分别主要在附睾和睾丸中表达。给无菌小鼠接种微生物会导致肠道中用于合成岩藻糖基化GA1的MFUT-II基因(Se基因)迅速增加。