title: Metal-ion induced conformational changes in alkaline phosphatase from E. coli assessed by limited proteolysis.
abstract: Alkaline phosphatase (AP) displays significant structural changes during metal-ion binding, supporting cooperative interactions between the subunits of the dimeric enzyme. Here, we present data on the dynamic properties of AP from E. coli, and characterize the structural changes that accompany variations in metal-ion content, combining limited proteolysis and MALDI-TOF mass spectrometry. Limited proteolysis revealed an internal cleavage site at Arg-293, reflecting a position of conformational flexibility supporting subunit communication essential for catalysis. A specific shielding of a region distant from the metal-binding site has been demonstrated, implying transmission of conformational changes, induced by metal-ion binding to the adjacent subunit, across the subunit interface.
authors.affiliation: Department of Chemistry, Faculty of Natural Sciences, Mathematics and Education, University of Split, N. Tesle 12, 21000 Split, Croatia.
authors.forename: V
authors.initials: V
authors.lastname: Bucević-Popović
authors.forename: M
authors.initials: M
authors.lastname: Pavela-Vrancic
authors.forename: R
authors.initials: R
authors.lastname: Dieckmann
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D001665:Binding Sites; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D007477:Ions / Q000378:metabolism / Q000494:pharmacology; D008274:Magnesium / Q000378:metabolism / Q000494:pharmacology; D008670:Metals / Q000378:metabolism* / Q000494:pharmacology; D011487:Protein Conformation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014357:Trypsin / Q000378:metabolism; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology
chemical_list: D029968:Escherichia coli Proteins; D007477:Ions; D008670:Metals; D000469:Alkaline Phosphatase; D014357:Trypsin; D008274:Magnesium; D015032:Zinc