Oligosaccharide recognition and binding to the carbohydrate binding module of AMP-activated protein kinase.
The AMP-activated protein kinase (AMPK) contains a carbohydrate-binding module (beta1-CBM) that is conserved from yeast to mammals. Beta1-CBM has been shown to localize AMPK to glycogen in intact cells and in vitro. Here we use Nuclear Magnetic Resonance spectroscopy to investigate oligosaccharide binding to 15N labelled beta1-CBM. We find that beta1-CBM shows greatest affinity to carbohydrates of greater than five glucose units joined via alpha,1-->4 glycosidic linkages with a single, but not multiple, glucose units in an alpha,1-->6 branch. The near identical chemical shift profile for all oligosaccharides whether cyclic or linear suggest a similar binding conformation and confirms the presence of a single carbohydrate-binding site.
Koay
Ann
A
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Victoria, Australia.
Rimmer
Kieran A
KA
Mertens
Haydyn D T
HD
Gooley
Paul R
PR
Stapleton
David
D
D055372:AMP-Activated Protein Kinases; D000818:Animals; D001665:Binding Sites; D009097:Multienzyme Complexes / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular; D009844:Oligosaccharides / Q000737:chemistry*; D011487:Protein Conformation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000737:chemistry*
D009097:Multienzyme Complexes; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; C055213:saccharide-binding proteins; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases