Structure-based discovery of a new affinity ligand to pancreatic alpha-amylase.
A ligand useful for affinity capture of porcine pancreatic alpha-amylase was found by virtual screening of the commercially available compound data base MDL Available Chemicals Directory. Hits from the virtual screening were investigated for binding by nuclear magnetic resonance (NMR) and surface plasmon resonance. Selected compounds were tested for inhibition of the enzyme using a NMR-based assay. One of the binders found was covalently coupled to a chromatographic resin and a column, packed with this resin, could retain alpha-amylase, which subsequently was eluted by introduction of the known inhibitor acarbose to the elution buffer.
Westerfors
Maria
M
Amersham Biosciences, Björkgatan 30, Uppsala, SE-75184, Sweden.
Tedebark
Ulf
U
Andersson
Hans O
HO
Ohrman
Sara
S
Choudhury
Devapriya
D
Ersoy
Oguz
O
Shinohara
Yasuro
Y
Axén
Andreas
A
Carredano
Enrique
E
Baumann
Herbert
H
D000818:Animals; D015374:Biosensing Techniques; D003198:Computer Simulation; D009682:Magnetic Resonance Spectroscopy; D008956:Models, Chemical; D008958:Models, Molecular; D010179:Pancreas / Q000201:enzymology*; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance; D013552:Swine; D000516:alpha-Amylases / Q000737:chemistry*
D000516:alpha-Amylases