Chandrasekaran E V, Jain R K, Larsen R D, Wlasichuk K, DiCioccio R A, Matta K L
Department of Gynecologic Oncology, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
Biochemistry. 1996 Jul 9;35(27):8925-33. doi: 10.1021/bi952194e.
Unique specificities of the cloned alpha 1,3-L-fucosyltransferases (FTs), FT III (Lewis type), FT IV (myeloid type), and FT V (plasma type), and the alpha 1,3-FTs of Colo 205 (colon carcinoma), HL 60 (myeloid), B142 (lymphoid), EKVX (lung carcinoma), and calf mesenteric lymph nodes (CMLN) were discerned with sulfated, sialylated, and/or fucosylated Gal beta 1,3/4GlcNAc beta-based acceptor moieties. (a) FT V was 1.0-, 20.8-, and 4.6-fold active in forming Lewis x, Lewis y, and 3'-alpha-galactosyl Lewis x, respectively. (b) FT III and FT V formed approximately 4-fold 3'-sulfo Lewis x, as compared to 3'-sialyl Lewis x. (c) FT IV showed great efficiency in forming 3'-sulfo Lewis x (249%) and Lewis x (345%) in mucin-type branched chains. (d) FT III, FT IV, and FT V formed 19%, 62%, and 47% 6-sulfo Lewis x as compared to Lewis x. (e) 6'-Sulfo Lewis x and 3'-sialyl-6'-sulfo Lewis x (GLYCAM ligand) were not synthesized from their immediate precursors by FT III, FT IV, or FT V. (f) FT III, FT IV, and FT V were 311%, 9%, and 188% active, respectively, with 2'-fucosyl lactose but were not active with 2'- fucosyl-6'-sulfo lactose. (g) FT III and FT V were 7.0- and 0.5-fold active in forming Lewis a as compared to Lewis x, whereas, FT IV was inactive. (h) FT III was -2.0-fold more active in forming 3'-alpha-galactosyl Lewis a than Lewis b. (i) FT III synthesized 6-sialyl Lewis a (40% efficiency as compared to Lewis a) from 6-sialyl type 1. (j) FT III did not act on 6'-sulfo or 6'-sialyl type 1 but was 106% and 22% active with 3'-sulfo and 6-sulfo type 1, respectively. (k) The Colo 205 FT activities with type 1 compounds almost paralleled that of FT III except for the low activity (9%) with Gal beta 1,3(NeuAc alpha 2, 6)GlcNAc beta-O-Bn, but with type 2 considerable differences between Colo 205 FT and FT III were noticed. (l) The alpha 1,3-FTs of CMLN, HL60, B142, and EKVX were 1.2-1.7 times active with Fuc alpha 1,2Gal beta 1,4GlcNAc beta- O-pNP and Gal alpha 1,3Gal beta 1,4 GlcNAc beta-O-Bn with respect to Gal beta 1,4GlcNAc beta-O-Al. (m) Both CMLN and HL60 FTs were 2-fold active with 3-sulfoGal beta 1,4GlcNAc in a mucin-type branch structure such as 3-sulfoGal beta 1,4GlcNAc beta 1,6(Gal beta 1,3)GalNAc alpha-O-Bn. (n) The 3'-sulfoLacNAc/acrylamide copolymer, either as an acceptor or as a competitive inhibitor, had the potential to distinguish myeloid type alpha 1,3-FT from the plasma type.
利用硫酸化、唾液酸化和/或岩藻糖基化的基于β1,3/4GlcNAc的Galβ受体部分，辨别出克隆的α1,3-L-岩藻糖基转移酶（FTs）、FT III（Lewis型）、FT IV（髓样型）和FT V（血浆型），以及来自Colo 205（结肠癌）、HL 60（髓样）、B142（淋巴样）、EKVX（肺癌）和小牛肠系膜淋巴结（CMLN）的α1,3-FTs的独特特异性。（a）FT V在形成Lewis x、Lewis y和3'-α-半乳糖基Lewis x时的活性分别为1.0倍、20.8倍和4.6倍。（b）与3'-唾液酸基Lewis x相比，FT III和FT V形成的3'-磺基Lewis x约为其4倍。（c）FT IV在粘蛋白型支链中形成3'-磺基Lewis x（249%）和Lewis x（345%）的效率很高。（d）与Lewis x相比，FT III、FT IV和FT V形成的6-磺基Lewis x分别为19%、62%和47%。（e）FT III、FT IV或FT V不能从其直接前体合成6'-磺基Lewis x和3'-唾液酸基-6'-磺基Lewis x（GLYCAM配体）。（f）FT III、FT IV和FT V对2'-岩藻糖基乳糖的活性分别为311%、9%和188%，但对2'-岩藻糖基-6'-磺基乳糖无活性。（g）与Lewis x相比，FT III和FT V在形成Lewis a时的活性分别为7.0倍和0.5倍，而FT IV无活性。（h）FT III在形成3'-α-半乳糖基Lewis a时比Lewis b的活性高-2.0倍。（i）FT III从6-唾液酸基1型合成6-唾液酸基Lewis a（与Lewis a相比效率为40%）。（j）FT III对6'-磺基或6'-唾液酸基1型无作用，但对3'-磺基和6-磺基1型的活性分别为106%和22%。（k）Colo 205 FT对1型化合物的活性几乎与FT III平行，只是对Galβ1,3（NeuAcα2,6）GlcNAcβ-O-Bn的活性较低（9%），但在2型化合物上，Colo 205 FT与FT III之间存在明显差异。（l）CMLN、HL60、B142和EKVX的α1,3-FTs对Fucα1,2Galβ1,4GlcNAcβ-O-pNP和Galα1,3Galβ1,4GlcNAcβ-O-Bn的活性相对于Galβ1,4GlcNAcβ-O-Al为1.2 - 1.7倍。（m）CMLN和HL60的FTs对粘蛋白型支链结构如3-磺基Galβ1,4GlcNAcβ1,6（Galβ1,3）GalNAcα-O-Bn中的3-磺基Galβ1,4GlcNAc的活性均为2倍。（n）3'-磺基乳糖胺/丙烯酰胺共聚物作为受体或竞争性抑制剂，有可能区分髓样型α1,3-FT和血浆型α1,3-FT。
