Inhibition of O-GlcNAcase Using a Potent and Cell-Permeable Inhibitor Does Not Induce Insulin Resistance in 3T3-L1 Adipocytes

TY - JOUR

T1 - Inhibition of O-GlcNAcase Using a Potent and Cell-Permeable Inhibitor Does Not Induce Insulin Resistance in 3T3-L1 Adipocytes

AU - Macauley, Matthew S.

AU - He, Yuan

AU - Gloster, Tracey M.

AU - Stubbs, Keith A.

AU - Davies, Gideon J.

AU - Vocadlo, David J.

PY - 2010/9/24

Y1 - 2010/9/24

N2 - To probe increased O-GlcNAc levels as an independent mechanism governing insulin resistance in 3T3-L1 adipocytes, a new class of O-GlcNAcase (OGA) inhibitor was studied. 6-Acetamido-6-deoxy-castanospermine (6-Ac-Cas) is a potent inhibitor of OGA. The structure of 6-Ac-Cas bound in the active site of an OGA homolog reveals structural features contributing to its potency. Treatment of 3T3-L1 adipocytes with 6-Ac-Cas increases O-GlcNAc levels in a dose-dependent manner. These increases in O-GlcNAc levels do not induce insulin resistance functionally, measured using a 2-deoxyglucose (2-DOG) uptake assay, or at the molecular level, determined by evaluating levels of phosphorylated IRS-1 and Akt. These results, and others described, provide a structural blueprint for improved inhibitors and collectively suggest that increased O-GlcNAc levels, brought about by inhibition of OGA, does not by itself cause insulin resistance in 3T3-L1 adipocytes.

AB - To probe increased O-GlcNAc levels as an independent mechanism governing insulin resistance in 3T3-L1 adipocytes, a new class of O-GlcNAcase (OGA) inhibitor was studied. 6-Acetamido-6-deoxy-castanospermine (6-Ac-Cas) is a potent inhibitor of OGA. The structure of 6-Ac-Cas bound in the active site of an OGA homolog reveals structural features contributing to its potency. Treatment of 3T3-L1 adipocytes with 6-Ac-Cas increases O-GlcNAc levels in a dose-dependent manner. These increases in O-GlcNAc levels do not induce insulin resistance functionally, measured using a 2-deoxyglucose (2-DOG) uptake assay, or at the molecular level, determined by evaluating levels of phosphorylated IRS-1 and Akt. These results, and others described, provide a structural blueprint for improved inhibitors and collectively suggest that increased O-GlcNAc levels, brought about by inhibition of OGA, does not by itself cause insulin resistance in 3T3-L1 adipocytes.

KW - BETA-N-ACETYLGLUCOSAMINIDASE

KW - PROTEIN MODIFICATION

KW - GLYCOSYL HYDROLASES

KW - NAG-THIAZOLINE

KW - IN-VIVO

KW - GLUCOSE

KW - PHOSPHORYLATION

KW - PUGNAC

KW - STATE

KW - GLCNACYLATION

UR - http://www.scopus.com/inward/record.url?scp=77956963665&partnerID=8YFLogxK

U2 - 10.1016/j.chembiol.2010.07.006

DO - 10.1016/j.chembiol.2010.07.006

M3 - Article

SN - 1074-5521

VL - 17

SP - 937

EP - 948

JO - Chemistry & Biology

JF - Chemistry & Biology

IS - 9

ER -