Toraya Tetsuo, Kuyama Atsushi, Tanaka Seiichi, Yamamoto Masatoyo, Ohmiya Tadamasa, Saito Yuri, Tanabe Tomoko
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka, Kita-ku, Okayama 700-8530, Japan.
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka, Kita-ku, Okayama 700-8530, Japan.
Biochem Biophys Res Commun. 2017 May 13;486(4):1055-1061. doi: 10.1016/j.bbrc.2017.03.161. Epub 2017 Mar 31.
Starfish oocytes are arrested at the prophase stage of the first meiotic division in the ovary. They resume meiosis by the stimulus of 1-methyladenine (1-MeAde), the maturation-inducing hormone for starfish oocytes. Putative 1-MeAde receptors have been suggested to be present on the oocyte surface, but not yet been characterized biochemically. As reported recently (T. Toraya, T. Kida, A. Kuyama, S. Matsuda, S. Tanaka, Y. Komatsu, T. Tsurukai, Biochem. Biophys. Res. Commun. 485 (2017) 41-46), it became possible to detect unknown 1-MeAde binders of starfish oocytes by immunophotoaffinity labeling, i.e., photoaffinity labeling combined with immunochemical detection. We designed and synthesized water-soluble and insoluble polymer-bound 1-MeAde derivatives. A water-soluble polymer-bound 1-MeAde derivative, in which 1-MeAde is bound to dextran through an N-substituent, triggered the germinal-vesicle breakdown toward follicle-free oocytes, dejellied oocytes, and denuded oocytes. This is consistent with the idea that putative 1-MeAde receptors are located on the cell surface of starfish oocytes. A water-insoluble polymer-bound 1-MeAde derivative, in which 1-MeAde is bound to Sepharose 4B through an N-substituent, served as an effective affinity adsorbent for the partial purification of a 1-MeAde binder with M of 47.5 K that might be a possible candidate of the maturation-inducing hormone receptors of starfish oocytes.
海星卵母细胞在卵巢中停滞于第一次减数分裂的前期阶段。它们通过1-甲基腺嘌呤(1-MeAde)的刺激恢复减数分裂,1-甲基腺嘌呤是海星卵母细胞的成熟诱导激素。推测1-MeAde受体存在于卵母细胞表面,但尚未进行生化特性鉴定。如最近报道(T. Toraya、T. Kida、A. Kuyama、S. Matsuda、S. Tanaka、Y. Komatsu、T. Tsurukai,《生物化学与生物物理研究通讯》485 (2017) 41 - 46),通过免疫光亲和标记,即光亲和标记与免疫化学检测相结合,能够检测海星卵母细胞中未知的1-MeAde结合物。我们设计并合成了水溶性和不溶性聚合物结合的1-MeAde衍生物。一种水溶性聚合物结合的1-MeAde衍生物,其中1-MeAde通过N-取代基与葡聚糖结合,可引发无滤泡卵母细胞、去胶膜卵母细胞和裸卵母细胞的生发泡破裂。这与推测的1-MeAde受体位于海星卵母细胞表面的观点一致。一种不溶性聚合物结合的1-MeAde衍生物,其中1-MeAde通过N-取代基与琼脂糖4B结合,可作为有效的亲和吸附剂,用于部分纯化一种分子量为47.5 K的1-MeAde结合物,该结合物可能是海星卵母细胞成熟诱导激素受体的候选物。