The fifth epidermal growth factor-like domain of thrombomodulin does not have an epidermal growth factor-like disulfide bonding pattern.
AuthorsWhite CE, Hunter MJ, Meininger DP, Garrod S, Komives EA
JournalProc Natl Acad Sci U S A
PubMed ID8816772
'The disulfide bonding pattern of the fourth and fifth epidermal growth factor (EGF)-like domains within the smallest active fragment of thrombomodulin have been determined. In previous work, this fragment was expressed and purified to homogeneity, and its cofactor activity, as measured by Kcat for thrombin activation of protein C, was ... More
Production of human tissue factor using the Pichia pastoris expression system.
AuthorsAustin AJ, Jones CE, Heeke GV
JournalProtein Expr Purif
PubMed ID9631526
Tissue factor plays an important role in the initiation of the blood coagulation cascade resulting in the formation of a fibrin clot. The extracellular domain of human tissue factor has been expressed in the protease-deficient strain of the methylotrophic yeast Pichia pastoris, SMD1168. Tissue factor was expressed with a human ... More
Identification and characterization of two alternative splice variants of human interleukin-2.
AuthorsTsytsikov VN, Yurovsky VV, Atamas SP, Alms WJ, White B
JournalJ Biol Chem
PubMed ID8798495
Our previous work showed that alternative splicing is used to make an inhibitory variant of human interleukin (IL)-4. Because of homology between IL-4 and IL-2 proteins and receptors, we tested whether alternative splicing is used to generate similar inhibitory variants of human IL-2. Messenger RNA from peripheral blood mononuclear cells ... More
Laccase from the white-rot fungus Trametes versicolor: cDNA cloning of lcc1 and expression in Pichia pastoris.
AuthorsJonsson LJ, Saloheimo M, Penttila M
JournalCurr Genet
PubMed ID9388299
A cDNA coding for laccase was isolated from the ligninolytic fungus Trametes versicolor by RNA-PCR. The cDNA corresponds to the gene lcc1, which encodes a laccase isoenzyme of 498 amino-acid residues preceded by a 22-residue signal peptide. The lcc1 cDNA was cloned into the vector pHIL-D2 for expression in Pichia ... More
The donor substrate specificity of the human beta 1,3-glucuronosyltransferase I toward UDP-glucuronic acid is determined by two crucial histidine and arginine residues.
The human beta1,3-glucuronosyltransferase I (GlcAT-I) plays a key role in proteoglycan biosynthesis by catalyzing the transfer of glucuronic acid onto the trisaccharide-protein linkage structure Galbeta1,3Galbeta1,4Xylbeta-O-Ser, a prerequisite step for polymerization of glycosaminoglycan chains. In this study, we identified His(308) and Arg(277) residues as essential determinants for the donor substrate (UDP-glucuronic ... More