5TD4

Starch binding sites on the Human pancreatic alpha amylase D300N variant complexed with an octaose substrate.

Summary for 5TD4

• Entry DOI: 10.2210/pdb5td4/pdb
• Related: 1CPU 4W93 4X9Y 5KEZ
• Related PRD ID: PRD_900001 PRD_900010 PRD_900035
• Descriptor: Pancreatic alpha-amylase, alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose, alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose, ... (7 entities in total)
• Functional Keywords: amylase, diabetes, obesity, glucosyl hydrolase, hydrolase
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 1
• Total formula weight: 59663.02

Authors

Caner, S.,Brayer, G.D. (deposition date: 2016-09-16, release date: 2016-11-02, Last modification date: 2024-10-30)

Primary citation

Zhang, X.,Caner, S.,Kwan, E.,Li, C.,Brayer, G.D.,Withers, S.G.
Evaluation of the Significance of Starch Surface Binding Sites on Human Pancreatic alpha-Amylase.
Biochemistry, 55:6000-6009, 2016

PubMed Abstract: Starch provides the major source of caloric intake in many diets. Cleavage of starch into malto-oligosaccharides in the gut is catalyzed by pancreatic α-amylase. These oligosaccharides are then further cleaved by gut wall α-glucosidases to release glucose, which is absorbed into the bloodstream. Potential surface binding sites for starch on the pancreatic amylase, distinct from the active site of the amylase, have been identified through X-ray crystallographic analyses. The role of these sites in the degradation of both starch granules and soluble starch was probed by the generation of a series of surface variants modified at each site to disrupt binding. Kinetic analysis of the binding and/or cleavage of substrates ranging from simple maltotriosides to soluble starch and insoluble starch granules has allowed evaluation of the potential role of each such surface site. In this way, two key surface binding sites, on the same face as the active site, are identified. One site, containing a pair of aromatic residues, is responsible for attachment to starch granules, while a second site featuring a tryptophan residue around which a malto-oligosaccharide wraps is shown to heavily influence soluble starch binding and hydrolysis. These studies provide insights into the mechanisms by which enzymes tackle the degradation of largely insoluble polymers and also present some new approaches to the interrogation of the binding sites involved.

PubMed: 27756128
DOI: 10.1021/acs.biochem.6b00992

Experimental method

X-RAY DIFFRACTION (2.3 Å)