'The synthesis of cyclic ADP-carbocyclic-ribose (cADPcR, 4) designed as a stable mimic of cyclic ADP-ribose (cADPR, 1), a Ca2+-mobilizing second messenger, was achieved using as the key step a condensation reaction with the phenylthiophosphate-type substrate 14 to form an intramolecular pyrophosphate linkage. The N-1-carbocyclic-ribosyladenosine derivative 16 was prepared via the ... More
Thr-161 Phosphorylation of Monomeric Cdc2. REGULATION BY PROTEIN PHOSPHATASE 2C IN XENOPUS OOCYTES.
'Fully grown Xenopus oocyte is arrested at prophase I of meiosis. Re-entry into meiosis depends on the activation of MPF (M-phase promoting factor or cyclin B.Cdc2 complex), triggered by progesterone. The prophase-arrested oocyte contains a store of Cdc2. Most of the protein is present as a monomer whereas a minor ... More
Neutrophil phospholipase D is activated by a membrane-associated Rho family small molecular weight GTP-binding protein.
AuthorsBowman EP, Uhlinger DJ, Lambeth JD
JournalJ Biol Chem
PubMed ID8408000
'Phospholipase D in human neutrophil lysates is activated by GTP gamma S (guanosine 5''-O-(3-thiotriphosphate)), implying the participation of a GTP-binding protein. Reconstitution of GTP gamma S-dependent activity requires protein factors in both the plasma membrane and the cytosol (Olson, S. C., Bowman, E. P., and Lambeth, J. D. (1991) J. ... More
The plasmid RK2 initiation protein binds to the origin of replication as a monomer.
AuthorsToukdarian AE, Helinski DR, Perri S
JournalJ Biol Chem
PubMed ID8636140
'The TrfA protein encoded by the broad host range bacterial plasmid RK2 specifically binds to eight direct repeats (iterons) present at the plasmid replication origin to initiate DNA replication. Purified TrfA protein is largely in the form of a dimer, and using a dimerization test system that involves the fusion ... More
Saccharomyces cerevisiae cytoplasmic tyrosyl-tRNA synthetase gene. Isolation by complementation of a mutant Escherichia coli suppressor tRNA defective in aminoacylation and sequence analysis.
AuthorsChow CM, RajBhandary UL
JournalJ Biol Chem
PubMed ID8509419
'Exploiting differences in tRNA recognition between prokaryotic and eukaryotic tyrosyl-tRNA synthetases (TyrRSs), we have isolated the gene for the cytoplasmic TyrRS of Saccharomyces cerevisiae by functional complementation in Escherichia coli of a mutant E. coli tRNA. The tRNA, derived from the E. coli initiator tRNA with changes to allow suppression ... More
A Calcium-Responsive Transcription Factor, CaRF, that Regulates Neuronal Activity-Dependent Expression of BDNF.
Authors Tao Xu; West Anne E; Chen Wen G; Corfas Gabriel; Greenberg Michael E;
JournalNeuron
PubMed ID11832226
'Transcription of the brain-derived neurotrophic factor (BDNF) gene is regulated in a calcium- and neuron-selective manner; however, the mechanisms that underlie this selectivity are not known. We have characterized a new calcium-response element, CaRE1, that is required for activity-dependent transcription of BDNF exon III and have cloned a transcription factor, ... More
Asef, a Link Between the Tumor Suppressor APC and G-Protein Signaling
The adenomatous polyposis coli gene (APC) is mutated in familial adenomatous polyposis and in sporadic colorectal tumors. Here the APC gene product is shown to bind through its armadillo repeat domain to a Rac-specific guanine nucleotide exchange factor (GEF), termed Asef. Endogenous APC colocalized with Asef in mouse colon epithelial ... More
ProNGF induces p75-mediated death of oligodendrocytes following spinal cord injury.
AuthorsBeattie MS, Harrington AW, Lee R, Kim JY, Boyce SL, Longo FM, Bresnahan JC, Hempstead BL, Yoon SO,
JournalNeuron
PubMed ID12408842
The neurotrophin receptor p75 is induced by various injuries to the nervous system, but its role after injury has remained unclear. Here, we report that p75 is required for the death of oligodendrocytes following spinal cord injury, and its action is mediated mainly by proNGF. Oligodendrocytes undergoing apoptosis expressed p75, ... More
Overexpression, characterization, and purification of a recombinant mouse immunophilin FKBP-52 and identification of an associated phosphoprotein.
To gain insight into the structure and function of the immunophilin FKBP-52, a mouse FKBP-52 was overexpressed in Spodoptera frugiperda insect cells (Sf9 cells) with the baculovirus expression system. The purification and characterization of the recombinant FKBP-52 (rFKBP-52) was facilitated by incorporating a histidine 6-mer domain at its N terminus. ... More
Tryptophan fluorescence reports nucleotide-induced conformational changes in a domain of the ArsA ATPase.
AuthorsZhou T, Rosen BP
JournalJ Biol Chem
PubMed ID9242630
The ars operon of plasmid R773 encodes an ATP-dependent extrusion pump for arsenite and antimonite in Escherichia coli. The ArsA ATPase is the catalytic subunit of the pump protein, with two nucleotide binding consensus sequences, one in the NH2-terminal half and one in the COOH- terminal half of the protein. ... More
Cooperative interaction between GATA-4 and GATA-6 regulates myocardial gene expression.
AuthorsCharron F, Paradis P, Bronchain O, Nemer G, Nemer M
JournalMol Cell Biol
PubMed ID10330176
Two members of the GATA family of transcription factors, GATA-4 and GATA-6, are expressed in the developing and postnatal myocardium and are equally potent transactivators of several cardiac promoters. However, several in vitro and in vivo lines of evidence suggest distinct roles for the two factors in the heart. Since ... More
Structure and ligand of a histone acetyltransferase bromodomain.
AuthorsDhalluin C, Carlson JE, Zeng L, He C, Aggarwal AK, Zhou MM
JournalNature
PubMed ID10365964
Histone acetylation is important in chromatin remodelling and gene activation. Nearly all known histone-acetyltransferase (HAT)-associated transcriptional co-activators contain bromodomains, which are approximately 110-amino-acid modules found in many chromatin-associated proteins. Despite the wide occurrence of these bromodomains, their three-dimensional structure and binding partners remain unknown. Here we report the solution structure ... More
Expression, purification, and characterization of gp160e, the soluble, trimeric ectodomain of the simian immunodeficiency virus envelope glycoprotein, gp160.
Authors Chen B; Zhou G; Kim M; Chishti Y; Hussey R E; Ely B; Skehel J J; Reinherz E L; Harrison S C; Wiley D C;
JournalJ Biol Chem
PubMed ID10944528
The envelope glycoprotein, gp160, of simian immunodeficiency virus (SIV) shares approximately 25% sequence identity with gp160 from the human immunodeficiency virus, type I, indicating a close structural similarity. As a result of binding to cell surface CD4 and co-receptor (e.g. CCR5 and CXCR4), both SIV and human immunodeficiency virus gp160 ... More
Identification and characterization of CIA/ASF1 as an interactor of bromodomains associated with TFIID.
General transcription initiation factor IID (TFIID) plays a central and critical role in transcription initiation from both naked and chromatin templates. Although interaction between several DNA-binding proteins and TFIID were identified and well characterized, functional linkage between TFIID and chromatin factors has remained to be elucidated. Here we show the ... More
The Golgi localization of phosphatidylinositol transfer protein beta requires the protein kinase C-dependent phosphorylation of serine 262 and is essential for maintaining plasma membrane sphingomyelin levels.
Authors van Tiel Claudia M; Westerman Jan; Paasman Marten A; Hoebens Martha M; Wirtz Karel W A; Snoek Gerry T;
JournalJ Biol Chem
PubMed ID11953429
Recombinant mouse phosphatidylinositol transfer protein (PI-TP)beta is a substrate for protein kinase C (PKC)-dependent phosphorylation in vitro. Based on site-directed mutagenesis and two-dimensional tryptic peptide mapping, Ser(262) was identified as the major site of phosphorylation and Ser(165) as a minor phosphorylation site. The phospholipid transfer activities of wild-type PI-TP beta ... More
Identification, molecular cloning, and characterization of a novel GABAA receptor-associated protein, GRIF-1.
AuthorsBeck M, Brickley K, Wilkinson HL, Sharma S, Smith M, Chazot PL, Pollard S, Stephenson FA.
JournalJ Biol Chem
PubMed ID12034717
A novel 913-amino acid protein, gamma-aminobutyric acid type A (GABA(A)) receptor interacting factor-1 (GRIF-1), has been cloned and identified as a GABA(A) receptor-associated protein by virtue of its specific interaction with the GABA(A) receptor beta 2 subunit intracellular loop in a yeast two-hybrid assay. GRIF-1 has no homology with proteins ... More
Characterization of three fragments that constitute the monomers of the human lysyl hydroxylase isoenzymes 1-3. The 30-kDa N-terminal fragment is not required for lysyl hydroxylase activity.
Lysyl hydroxylase (LH) catalyzes the formation of hydroxylysine in collagens; three human isoenzymes have been cloned so far. We report here on the purification of all three recombinant isoenzymes to homogeneity from the medium of cultured insect cells, and we demonstrate that they are all homodimers. Limited proteolysis experiments identified ... More