caa a22 4500 397557698 CHVBK 20180308164807.0 cr unu---uuuuu 161202e199605 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021335 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021335 Unique Recognition of Activin and Inhibin by Polyclonal Antibodies to Inhibin Subunits [Elektronische Daten] [Masayuki Funaba, Takuya Murata, Hisako Fujimura, Eri Murata, Matanobu Abe, Michio Takahashi, Kunio Torii] Inhibin-A is a glycoprotein composed of an a subunit containing a glycosylation site and a βA subunit, whereas activin-A is a homodimer of two inhibin βA subunits. We examined the recognition of activin-A and inhibin-A by several antisera to the α or βA; subunit, and factors affecting the recognition. A total of six polyclonal antibodies to inhibin subunits, i.e., two antisera to a peptide fragment of the α subunit [α(l-19) and α(lα26)], and four antisera to the βA subunit [βA(l-10), βA(70-79), βA(87-99), and βA(94-105)], was generated. On Western blot analysis, the anti-βA(87-99) and βA(94-105) sera recognized recombinant human activin-A but not inhibin-A under non-reducing conditions. When inhibin-A was deglycosylated with N-glycosidase-F, inhibin-A could be recognized by the anti-βA(87-99) and βA(94-105) sera. In addition, when activin-A bound to a nitrocellulose membrane was p re-incubated with recombinant human follistatin, the recognition of activin-A by the anti-βA(87-99) and βA(94-105) sera was decreased. These results suggested that the lower affinity of follistatin to inhibin-A than to activin-A might be likely explained as reflecting a site associated with the glycosylation of inhibin-A. However, the exposure of amino acids 87-105 of the inhibin 0A subunit on the molecular surface through deglycosylation did not increase the affinity of inhibin-A for follistatin but rather resulted in poor binding with follistatin. The present data suggest that (1) amino acids 87-105 of the inhibin/activin βA subunit are located on the molecular surface, although this region of inhibin-A is concealed by the carbohydrate chain of the α subunit, (2) the region responsible for follistatin binding within the activin βA subunit is spanned by amino acids 87-105, and (3) the mode of binding of inhibin-A to follistatin is quite different from that of activin-A to follistatin, and the former may be influenced by glycosylation. © by the Journal of Biochemistry Regular Papers nationallicence activin nationallicence follistatin nationallicence inhibin nationallicence polyclonal antibodies nationallicence recognition nationallicence Funaba Masayuki Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut Murata Takuya Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut Fujimura Hisako Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut Murata Eri Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut Abe Matanobu School of Veterinary Medicine, Azabu University Sagamihara 229 aut Takahashi Michio Department of Veterinary Physiology, The University of Tokyo 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113 aut Torii Kunio Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut The Journal of Biochemistry Oxford University Press 119/5(1996-05), 953-960 0021-924X 119:5<953 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021335 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021335 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Funaba Masayuki Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut NATIONALLICENCE 700 1- Murata Takuya Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut NATIONALLICENCE 700 1- Fujimura Hisako Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut NATIONALLICENCE 700 1- Murata Eri Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut NATIONALLICENCE 700 1- Abe Matanobu School of Veterinary Medicine, Azabu University Sagamihara 229 aut NATIONALLICENCE 700 1- Takahashi Michio Department of Veterinary Physiology, The University of Tokyo 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113 aut NATIONALLICENCE 700 1- Torii Kunio Torii Nutrient-Stasis Project, ERATO, R & D Corp. of Japan Yokohama 221 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/5(1996-05), 953-960 0021-924X 119:5<953 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford