Biomedical subjects

Seppo Lapinjoki

Publications and source records attributed to Seppo Lapinjoki.

3 recordsLinked to original sources

PubMed2006

Ribokinase from E. coli: expression, purification, and substrate specificity.

Ribokinase (RK) was expressed in the Escherichia coli ER2566 cells harboring the constructed expression plasmid encompassing the rbsK gene, encoding ribokinase. The recombinant enzyme was purified from sonicated cells by double chromatography to afford a preparation that was ca. 90% pure and had specific activity of 75 micromol/min mg protein. Catalytic activity of RK: (i) is strongly dependent on the presence of monovalent cations (potassium>>>ammonium>cesium), and (ii) is cooperatively enhanced by divalent magnesium and manganese ions. Besides D-ribose and 2-deoxy-D-ribose, RK was found to catalyze the 5-O-phosphorylation of D-arabinose, D-xylose, and D-fructose in the presence of ATP, and potassium and magnesium ions; L-ribose and L-arabinose are not substrates for the recombinant enzyme. A new radiochemical method for monitoring the formation of D-pentofuranose-5-[32P]phosphates in the presence of [gamma-32P]ATP and RK is reported.

Binding Sites↗

PubMed2005

Human and bovine milk oligosaccharides inhibit Neisseria meningitidis pili attachment in vitro.

Milk oligosaccharides have been shown to interfere with adhesion of many pathogens to host mucosal surfaces. Characterization of the adhesion mechanisms of the bacteria to host cell surface is needed to develop novel functional food, infant formulas, and anti-infective drugs. Adhesion of Neisseria meningitidis, a human specific pathogen causing meningitis and septicemia, is not completely understood but is mediated by type IV pili. Here, we developed a microtiter well pili binding assay to investigate the binding activities of N. meningitidis isolated type IV pili to different glycoproteins. Pili binding activities to bovine thyroglobulin and human salivary agglutinin but not to chicken ovalbumin were present. Inhibition of these binding activities was demonstrated by fractionated human or bovine milk oligosaccharides. The binding of neisserial pili to bovine thyroglobulin was most effective and was clearly inhibited by human milk neutral or bovine milk acidic oligosaccharides.

Animals↗

PubMed2002

High-performance liquid chromatography--mass spectrometry and electron-capture dissociation tandem mass spectrometry of osteocalcin. Determination of gamma-carboxyglutamic acid residues.

Two mass spectrometry methods, high-performance liquid chromatography combined on-line with electrospray ionization mass spectrometry (HPLC-ESI-MS) and electron-capture (EC) dissociation tandem mass spectrometry (MS-MS), were applied for structural analysis of bovine and human osteocalcins. Osteocalcin contains gamma-carboxyglutamic acid (Gla) residues, which bind metal ions, among its amino acids. Ethylenediaminetetraacetic acid (EDTA) was added to all samples in order to chelate bound metal ions. After elimination of interfering metal ions MS spectra became uncomplicated to interpret. EDTA is incompatible with ESI and it was removed from samples using either on-line HPLC or micropurification method. The number of Gla residues varies in osteocalcin. These subforms, which contain different amounts of Gla residues, were separated using the HPLC-ESI-MS method. In order to determine locations of Gla residues in human osteocalcin, which contained two Gla residues, dissociation MS-MS method was successfully applied.

1-Carboxyglutamic Acid↗