A glycosidase antibody elicited against a chair-like transition state analog by in vitro immunization.
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 beta-D-glucopyranoside 3 with a rate enhancement (kcat/kuncat) of 10(5) M 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.
Yu
J
J
Division of Applied Science, The Korea Institute of Science and Technology, P.O. Box 131, Cheongryang, Seoul 131-791 Korea.
Choi
S Y
SY
Moon
K D
KD
Chung
H H
HH
Youn
H J
HJ
Jeong
S
S
Park
H
H
Schultz
P G
PG
D017485:1-Deoxynojirimycin / Q000031:analogs & derivatives; D017151:Antibodies, Catalytic / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000276:immunology; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000276:immunology; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors*; D006027:Glycosides / Q000378:metabolism*; D006241:Haptens / Q000276:immunology
D017151:Antibodies, Catalytic; D004791:Enzyme Inhibitors; D006027:Glycosides; D006241:Haptens; D017485:1-Deoxynojirimycin; D006026:Glycoside Hydrolases; D005944:Glucosamine; C013626:nojirimycin