A Glycosidase Antibody Elicited against a Chair-Like Transition State Analog by in vitro Immunization

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 95; no. 6; pp. 2880 - 2884
• Main Authors: Yu, Jaehoon, Choi, So Young, Moon, Kyung-Duk, Chung, Hyun-Ho, Youn, Hyun Joo, Jeong, Sunjoo, Park, Hokoon, Schultz, Peter G.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 17.03.1998; National Acad Sciences; National Academy of Sciences; The National Academy of Sciences
• Subjects: 1-Deoxynojirimycin - analogs & derivatives; Active sites; Antibodies; Antibodies, Catalytic - metabolism; Biochemistry; Biological Sciences; Carboxylates; Catalytic activity; Enzyme Inhibitors - immunology; Glucosamine - analogs & derivatives; Glucosamine - chemistry; Glucosamine - immunology; Glycoside Hydrolases - antagonists & inhibitors; Glycosides - metabolism; Haptens; Haptens - immunology; Hybridomas; Hydrolysis; Immune system; Immunization; Ions
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.95.6.2880

Antibodies were generated against the positively charged chair-like glycosidase inhibitor nojirimycin by in vitro immunization. A number of catalytic antibodies were isolated, one of which catalyzes the hydrolysis of p-nitrophenyl β -D-glucopyranoside 3 with a rate enhancement (kcat/kuncat) of 105M over the HOAC-catalyzed reaction. The antibody discriminates modifications in the pyranoside ring of substrate 3 at the C2, C4, and the anomeric positions. The pH dependence of the reaction and chemical modification studies suggest the presence of an active-site Asp or Glu residue that may function as a general acid. This study further defines those requirements necessary to generate antibodies that efficiently cleave glycosidic bonds.