title: Structure of human salivary alpha-amylase crystallized in a C-centered monoclinic space group.
abstract: Human salivary alpha-amylase (HSA) is a major secretory protein component of saliva and has important biological functions, including the initial digestion of starch. HSA acts as a monomer and mediates the hydrolysis of alpha-1,4-glucosidic linkages in oligosaccharides. To date, all published crystal structures of HSA have been crystallized as monomers in space group P2(1)2(1)2(1). Here, the serendipitous purification, crystallization and ultimate structure determination of a HSA non-crystallographic symmetry (NCS) dimer, while attempting to purify human carbonic anhydrase VI (HCA VI) from saliva using an affinity resin for alpha-class carbonic anhydrases, is presented. On further investigation, it was shown that HSA could only be copurified using the affinity resin in the presence of HCA VI which is glycosylated and not the non-glycosylated HCA II. The identification of the HSA crystals was carried out by peptide mapping and mass spectrometry. HSA was shown to have crystallized as an NCS dimer in space group C2, with unit-cell parameters a = 150.9, b = 72.3, c = 91.3 A, beta = 102.8 degrees. The NCS dimer crystal structure is reported to 3.0 A resolution, with a refined Rcryst of 0.228. The structure is compared with the previously reported P2(1)2(1)2(1) monomer structures and the crystal packing and dimer interface are discussed.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32601, USA.
authors.forename: S Zoë
authors.initials: SZ
authors.lastname: Fisher
authors.forename: Lakshmanan
authors.initials: L
authors.lastname: Govindasamy
authors.forename: Chingkuang
authors.initials: C
authors.lastname: Tu
authors.forename: Mavis
authors.initials: M
authors.lastname: Agbandje-McKenna
authors.forename: David N
authors.initials: DN
authors.lastname: Silverman
authors.forename: Hannu J
authors.initials: HJ
authors.lastname: Rajaniemi
authors.forename: Robert
authors.initials: R
authors.lastname: McKenna
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D019281:Dimerization; D006031:Glycosylation; D006801:Humans; D010449:Peptide Mapping; D012463:Saliva / Q000201:enzymology*; D000516:alpha-Amylases / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D000516:alpha-Amylases