Rhodopsin kinase: expression in mammalian cells and a two-step purification.
AuthorsBruel C, Cha K, Reeves PJ, Getmanova E, Khorana HG
JournalProc Natl Acad Sci U S A
PubMed ID10737781
'A suitable system for expression of the rhodopsin kinase (RK) gene and its mutants is needed for structure-function studies of RK. Previously, investigation of the baculovirus system showed satisfactory production of RK, but posttranslational isoprenylation was deficient. We now report on a comparative study of expression of the RK gene ... More
Biochemical and antigenic characterization of a new dipeptidyl- peptidase isolated from Aspergillus fumigatus.
AuthorsBeauvais A, Monod M, Debeaupuis JP, Diaquin M, Kobayashi H, Latge JP
JournalJ Biol Chem
PubMed ID9045640
A novel dipeptidyl-peptidase (DPP V) was purified from the culture medium of Aspergillus fumigatus. This is the first report of a secreted dipeptidyl-peptidase. The enzyme had a molecular mass of 88 kDa and contained approximately 9 kDa of N-linked carbohydrate. The expression and secretion of dipeptidyl-peptidase varied with the growth ... More
Expression of recombinant HLA-DR2 molecules. Replacement of the hydrophobic transmembrane region by a leucine zipper dimerization motif allows the assembly and secretion of soluble DR alpha beta heterodimers.
AuthorsKalandadze A, Galleno M, Foncerrada L, Strominger JL, Wucherpfennig KW
JournalJ Biol Chem
PubMed ID8702739
Major histocompatibility complex (MHC) class II molecules are membrane-anchored heterodimers that present peptides on the surface of antigen presenting cells to T cells. Soluble HLA-DR2 molecules were expressed for structural and functional characterization of the MHC/peptide/T cell receptor recognition unit. The alpha and beta chains of DR2 (encoded by the ... More
Potency and selectivity of the cathepsin L propeptide as an inhibitor of cysteine proteases.
AuthorsCarmona E, Dufour E, Plouffe C, Takebe S, Mason P, Mort JS, Menard R
JournalBiochemistry
PubMed ID8679567
The cathepsin L propeptide (phcl-2) was expressed in Saccharomyces cerevisiae using a human procathepsin L/alpha-factor fusion construct containing a stop codon at position -1 (the C-terminal amino acid of the proregion). Since the yield after purification was very low, the cathepsin L propeptide was also obtained by an alternate procedure ... More
Manipulating monomer-dimer equilibrium of bovine Beta -lactoglobulin by amino acid substitution.
Authors Sakurai Kazumasa; Goto Yuji;
JournalJ Biol Chem
PubMed ID12006601
Bovine beta-lactoglobulin, a major protein in cow's milk composed of nine beta-strands (betaA-betaI) and one alpha-helix, exists as a dimer at neutral pH while it dissociates to a native monomer below pH 3.0. It is assumed that the intermolecular beta-sheet formed between I-strands and salt bridges at AB-loops play important ... More