'OX40 is a member of the tumor necrosis factor (TNF) receptor superfamily and known to be an important costimulatory molecule expressed on activated T cells. To investigate the role of costimulation of OX40 in human immunodeficiency virus type 1 (HIV-1) infection by its natural ligand, gp34, the OX40-transfected ACH-2 cell ... More
Kinesin Superfamily Motor Protein KIF17 and mLin-10 in NMDA Receptor-Containing Vesicle Transport
'Experiments with vesicles containing N-methyl-D-aspartate (NMDA)receptor 2B (NR2B subunit) show that they are transported alongmicrotubules by KIF17, a neuron-specific molecular motor in neuronaldendrites. Selective transport is accomplished by direct interaction of theKIF17 tail with a PDZ domain of mLin-10 (Mint1/X11), which is aconstituent of a large protein complex including mLin-2 ... More
Alphaherpesvirus origin-binding protein homolog encoded by human herpesvirus 6B, a betaherpesvirus, binds to nucleotide sequences that are similar to ori regions of alphaherpesviruses.
AuthorsInoue N, Dambaugh TR, Rapp JC, Pellett PE
JournalJ Virol
PubMed ID8207791
'We previously identified a human herpesvirus 6B (HHV-6B) homolog of the alphaherpesvirus origin-binding protein (OBP), exemplified by the herpes simplex virus type 1 UL9 gene product. This finding is of particular interest because HHV-6B is otherwise more closely related to members of the betaherpesvirus subfamily. The prototypic betaherpesvirus, human cytomegalovirus, ... More
Interaction of human nuclear topoisomerase I with guanosine quartet-forming and guanosine-rich single-stranded DNA and RNA oligonucleotides.
'Human nuclear DNA topoisomerase I (top1) plays a crucial role in DNA replication, transcription, and chromosome condensation. In this study, we show that intra- and intermolecular guanosine quartets (G-quartets) can inhibit top1-mediated DNA cleavage at a high affinity site. Top1-mediated DNA cleavage was also inhibited by a 16-mer single-stranded oligodeoxynucleotide ... More
Purification and characterization of human DNA damage checkpoint Rad complexes.
Authors Lindsey-Boltz L A; Bermudez V P; Hurwitz J; Sancar A;
JournalProc Natl Acad Sci U S A
PubMed ID11572977
'Checkpoint Rad proteins function early in the DNA damage checkpoint signaling cascade to arrest cell cycle progression in response to DNA damage. This checkpoint ensures the transmission of an intact genetic complement to daughter cells. To learn about the damage sensor function of the human checkpoint Rad proteins, we purified ... More
Expression and characterization of functionally active recombinant perforin produced in insect cells.
AuthorsLiu CC, Persechini PM, Young JD
JournalJ Immunol
PubMed ID8617952
'A key cytolytic mediator used by killer lymphocytes, perforin (also known as pore-forming protein or cytolysin), has been shown to be capable of undergoing polymerization to form pores in cell membranes and cause osmotic lysis of target cells. Although perforin has been purified from killer lymphocytes and the coding gene ... More
A dominant-negative mutant of human DNA helicase B blocks the onset of chromosomal DNA replication.
Authors Taneja Poonam; Gu Jinming; Peng Rui; Carrick Ryan; Uchiumi Fumiaki; Ott Robert D; Gustafson Eric; Podust Vladimir N; Fanning Ellen;
JournalJ Biol Chem
PubMed ID12181327
'A cDNA encoding a human ortholog of mouse DNA helicase B, which may play a role in DNA replication, has been cloned and expressed as a recombinant protein. The predicted human DNA helicase B (HDHB) protein contains conserved helicase motifs (superfamily 1) that are strikingly similar to those of bacterial ... More
Alternate interactions define the binding of peptides to the MHC molecule IA(b).
Authors Liu Xinqi; Dai Shaodong; Crawford Frances; Fruge Rachel; Marrack Philippa; Kappler John;
JournalProc Natl Acad Sci U S A
PubMed ID12084926
'We have solved the crystal structure of the MHCII molecule, IA(b), containing an antigenic variant of the major IA(b)-binding peptide derived from the MHCII IEalpha chain. The four MHC pockets at p1, p4, p6, and p9 that usually bind peptide side chains are largely empty because of alanines in the ... More
Baculovirus expression and purification of rat 10- formyltetrahydrofolate dehydrogenase.
AuthorsKrupenko SA, Horstman DA, Wagner C, Cook RJ
JournalProtein Expr Purif
PubMed ID8527931
'The liver cytosolic enzyme, 10-formyltetrahydrofolate dehydrogenase (10- FTHFDH) (EC 1.5.1.6) catalyzes two reactions: the NADP(+)-dependent oxidation of 10-formyltetrahydrofolate to tetrahydrofolate and CO2 and the NADP(+)-independent hydrolysis of 10-formyltetrahydrofolate to tetrahydrofolate and formate. It exists as a tetramer of 99-kDa subunits in rat liver. We expressed rat liver cDNA encoding 10-FTHFDH ... More
A Novel Type I Cytokine Receptor Is Expressed on Monocytes, Signals Proliferation, and Activates STAT-3 and STAT-5.
Authors Ghilardi Nico; Li Ji; Hongo Jo-Anne; Yi Sothy; Gurney Austin; De Sauvage Frederic J;
JournalJ Biol Chem
PubMed ID11877449
'Here we report the cloning of a novel type I cytokine receptor, gp130-like monocyte receptor (GLM-R), with homology to the interleukin-6 receptor signal transducing chain, gp130, and granulocyte colony-stimulating factor receptor. Human and murine GLM-R cDNAs encode open reading frames of 732 and 716 amino acids, respectively, and the corresponding ... More
Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment.
AuthorsWickham TJ, Mathias P, Cheresh DA, Nemerow GR
JournalCell
PubMed ID8477447
'Adenovirus contains a heterodimeric protein complex consisting of 186 kd fiber protein that mediates high affinity virus attachment to cells and a 400 kd pentavalent subunit (penton base) that contains five Arg- Gly-Asp sequences, implying a role for integrins in adenovirus infection. We demonstrate that the vitro-nectin-binding integrins alpha v ... More
The exoskeleton collagens in Caenorhabditis elegans are modified by prolyl 4-hydroxylases with unique combinations of subunits.
'The collagen prolyl 4-hydroxylases (P4Hs, EC ) play a critical role in the synthesis of the extracellular matrix. The enzymes characterized from vertebrates and Drosophila are alpha(2)beta(2) tetramers, in which protein disulfide isomerase (PDI) serves as the beta subunit. Two conserved alpha subunit isoforms, PHY-1 and PHY-2, have been identified ... More
Detection of antigen-specific T cells with multivalent soluble class II MHC covalent peptide complexes.
AuthorsCrawford F, Kozono H, White J, Marrack P, Kappler J
JournalImmunity
PubMed ID9655481
Multimeric soluble MHC class II molecules stably occupied with covalently attached peptides bind with appropriate specificity to T cell hybridomas and T cells from T cell receptor transgenic mice. There is a direct correlation between soluble T cell receptor affinity for monomeric MHC/peptide and level of binding of multimeric MHC/peptide ... More
H2-M (HLA-DM in humans) resides in an acidic endosomal compartment, where it facilitates the loading of antigenic peptides into the peptide- binding groove of class II MHC. The crystal structure of a soluble form of H2-M has been solved to 3.1 A resolution, revealing a heterodimer with structural similarities to ... More
Coordinate binding of ATP and origin DNA regulates the ATPase activity of the origin recognition complex.
AuthorsKlemm RD, Austin RJ, Bell SP
JournalCell
PubMed ID9038340
The Origin Recognition Complex (ORC) is a six-protein assembly that specifies the sites of DNA replication initiation in S. cerevisiae. Origin recognition by ORC requires ATP. Here, we demonstrate that two subunits, Orc1p and Orc5p, bind ATP and that Orc1p also hydrolyzes ATP. ATP binding and hydrolysis by Orc1p are ... More
Human Tra2 proteins are sequence-specific activators of pre-mRNA splicing.
AuthorsTacke R, Tohyama M, Ogawa S, Manley JL
JournalCell
PubMed ID9546399
The RNA-binding protein Tra2 is an important regulator of sex determination in Drosophila. Recently, two mammalian Tra2 homologs of unknown function have been described. Here, we show that human Tra2 proteins are present in HeLa cell nuclear extracts and that they bind efficiently and specifically to a previously characterized pre-mRNA ... More
Conversion of proepithelin to epithelins: roles of SLPI and elastase in host defense and wound repair.
Increased leukocyte elastase activity in mice lacking secretory leukocyte protease inhibitor (SLPI) leads to impaired wound healing due to enhanced activity of TGFbeta and perhaps additional mechanisms. Proepithelin (PEPI), an epithelial growth factor, can be converted to epithelins (EPIs) in vivo by unknown mechanisms with unknown consequences. We found that ... More
Endothelial induction of fgl2 contributes to thrombosis during acute vascular xenograft rejection.
AuthorsGhanekar A, Mendicino M, Liu H, He W, Liu M, Zhong R, Phillips MJ, Levy GA, Grant DR,
JournalJ Immunol
PubMed ID15100314
Thrombosis is a prominent feature of acute vascular rejection (AVR), the current barrier to survival of pig-to-primate xenografts. Fibrinogen-like protein 2 (fgl2/fibroleukin) is an inducible prothrombinase that plays an important role in the pathogenesis of fibrin deposition during viral hepatitis and cytokine-induced fetal loss. We hypothesized that induction of fgl2 ... More
Recombinant Dicer efficiently converts large dsRNAs into siRNAs suitable for gene silencing.
AuthorsMyers JW, Jones JT, Meyer T, Ferrell JE,
JournalNat Biotechnol
PubMed ID12592410
RNA interference (RNAi) is a powerful method for specifically silencing gene expression in diverse cell types. RNAi is mediated by approximately 21-nucleotide small interfering RNAs (siRNAs), which are produced from larger double-stranded RNAs (dsRNAs) in vivo through the action of Dicer, an RNase III-family enzyme. Transfecting cells with siRNAs rather ... More
Influence of the NH2-terminal amino acid of the T cell receptor alpha chain on major histocompatibility complex (MHC) class II + peptide recognition.
AuthorsSeibel JL, Wilson N, Kozono H, Marrack P, Kappler JW
JournalJ Exp Med
PubMed ID9166421
The alpha/beta T cell receptor (TCR) recognizes peptide fragments bound in the groove of major histocompatibility complex (MHC) molecules. We modified the TCR alpha chain from a mouse T cell hybridoma and tested its ability to reconstitute TCR expression and function in an alpha chain-deficient variant of the hybridoma. The ... More
Expression, purification, and properties of the aldehyde dehydrogenase homologous carboxyl-terminal domain of rat 10-formyltetrahydrofolate dehydrogenase.
AuthorsKrupenko SA, Wagner C, Cook RJ
JournalJ Biol Chem
PubMed ID9092577
The liver cytosolic enzyme, 10-formyltetrahydrofolate dehydrogenase (FDH) (EC 1.5.1.6) catalyzes two reactions: the NADP+-dependent oxidation of 10-formyltetrahydrofolate to tetrahydrofolate and CO2 and the NADP+-independent hydrolysis of 10-formyltetrahydrofolate to tetrahydrofolate and formate. The COOH-terminal domain of the enzyme (residues 420-902) is about 48% identical to a family of NAD-dependent aldehyde dehydrogenases ... More
The product of a novel growth factor-activated gene, fic, is a biologically active C-C-type cytokine.
AuthorsHeinrich JN, Ryseck RP, Macdonald-Bravo H, Bravo R
JournalMol Cell Biol
PubMed ID8455595
We have characterized a new member of the superfamily of proinflammatory peptides encoded by a growth factor-inducible gene, fic, previously isolated by differential screening of a cDNA library of mRNA from serum-stimulated NIH 3T3 cells. Immunoprecipitation analyses showed that the protein was rapidly induced following serum stimulation and secreted unglycosylated ... More
The significance of tetramerization in promoter recruitment by Stat5.
AuthorsJohn S, Vinkemeier U, Soldaini E, Darnell JE Jr, Leonard WJ
JournalMol Cell Biol
PubMed ID10022878
Stat5a and Stat5b are rapidly activated by a wide range of cytokines and growth factors, including interleukin-2 (IL-2). We have previously shown that these signal transducers and activators of transcription (STAT proteins) are key regulatory proteins that bind to two tandem gamma interferon-activated site (GAS) motifs within an IL-2 response ... More
Dissection of CR1, factor H, membrane cofactor protein, and factor B binding and functional sites in the third complement component.
Previous studies have suggested that the residues 727-768 of human (Hu) C3 contain the binding sites for CR1, factor H, and factor B. Here, we have (1) characterized further some of the C3 structural requirements for its binding to CR1, H, and B, (2) investigated the functions associated with these ... More
Heparin promotes proteolytic inactivation by thrombin of a reactive site mutant (L444R) of recombinant heparin cofactor II.
AuthorsCiaccia AV, Willemze AJ, Church FC
JournalJ Biol Chem
PubMed ID8995378
A heparin cofactor II (HCII) mutant with an Arg substituted for Leu444 at the P1 position (L444R-rHCII) was previously found to have altered proteinase specificity (Derechin, V. M., Blinder, M. A., and Tollefsen, D. M. (1990) J. Biol. Chem. 265, 5623-5628). The present study characterizes the effect of glycosaminoglycans on ... More
Kit receptor dimerization is driven by bivalent binding of stem cell factor.
AuthorsLemmon MA, Pinchasi D, Zhou M, Lax I, Schlessinger J
JournalJ Biol Chem
PubMed ID9045650
Most growth factors and cytokines activate their receptors by inducing dimerization upon binding. We have studied binding of the dimeric cytokine stem cell factor (SCF) to the extracellular domain of its receptor Kit, which is a receptor tyrosine kinase similar to the receptors for platelet-derived growth factor and colony-stimulating factor-1. ... More
Molecular characterization of major cat allergen Fel d 1: expression of heterodimer by use of a baculovirus expression system.
Fel d 1 is a major cat allergen inducing allergic rhinitis and asthma in sensitized individuals. It has a more complex structure when compared with other allergens and therefore expression of recombinant Fel d 1 has been considered a challenge. The present study shows for the first time that a ... More
Functional analysis of mutant and wild-type Drosophila origin recognition complex.
Authors Chesnokov I; Remus D; Botchan M;
JournalProc Natl Acad Sci U S A
PubMed ID11593009
The origin recognition complex (ORC) is the DNA replication initiator protein in eukaryotes. We have reconstituted a functional recombinant Drosophila ORC and compared activities of the wild-type and several mutant ORC variants. Drosophila ORC is an ATPase, and our studies show that the ORC1 subunit is essential for ATP hydrolysis ... More
Asp-225 and glu-375 in autocatalytic attachment of the prosthetic heme group of lactoperoxidase.
Authors Colas Christophe; Kuo Jane M; Ortiz de Montellano Paul R;
JournalJ Biol Chem
PubMed ID11756449
The heme in lactoperoxidase is attached to the protein by ester bonds between the heme 1- and 5-methyl groups and Glu-375 and Asp-275, respectively. To investigate the cross-linking process, we have examined the D225E, E375D, and D225E/E375D mutants of bovine lactoperoxidase. The heme in the E375D mutant is only partially ... More
The type XIII collagen ectodomain is a 150-nm rod and capable of binding to fibronectin, nidogen-2, perlecan, and heparin.
Type XIII collagen consists of a short N-terminal intracellular domain, a transmembrane domain, and a collagenous ectodomain, and it is found at many sites of cell adhesion. We report on the characterization of recombinant type XIII collagen. The shed ectodomain was purified from insect cell culture medium and shown to ... More
Functional conservation of subfamilies of putative UDP-N-acetylgalactosamine:polypeptide N-acetylgalactosaminyltransferases in Drosophila, Caenorhabditis elegans, and mammals. One subfamily composed of l(2)35Aa is essentail in Drosophila.
Authors Schwientek Tilo; Bennett Eric P; Flores Carlos; Thacker John; Hollmann Martin; Reis Celso A; Behrens Jane; Mandel Ulla; Keck Birgit; Schäfer Mireille A; Haselmann Kim; Zubarev Roman; Roepstorff Peter; Burchell Joy M; Taylor-Papadimitriou Joyce; Hollingsworth Michael A; Clausen Henrik;
JournalJ Biol Chem
PubMed ID11925450
The completed fruit fly genome was found to contain up to 15 putative UDP-N-acetyl-alpha-d-galactosamine:polypeptide N-acetylgalactosaminyltransferase (GalNAc-transferase) genes. Phylogenetic analysis of the putative catalytic domains of the large GalNAc-transferase enzyme families of Drosophila melanogaster (13 available), Caenorhabditis elegans (9 genes), and mammals (12 genes) indicated that distinct subfamilies of orthologous genes ... 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
Punctin, a novel ADAMTS-like molecule, ADAMTSL-1, in extracellular matrix.
Authors Hirohata Satoshi; Wang Lauren W; Miyagi Masaru; Yan Lin; Seldin Michael F; Keene Douglas R; Crabb John W; Apte Suneel S;
JournalJ Biol Chem
PubMed ID11805097
Punctin (ADAMTSL-1) is a secreted molecule resembling members of the ADAMTS family of proteases. Punctin lacks the pro-metalloprotease and the disintegrin-like domain typical of this family but contains other ADAMTS domains in precise order including four thrombospondin type I repeats. Punctin is the product of a distinct gene on human ... More
The lymphocyte metalloprotease MDC-L (ADAM 28) is a ligand for the integrin alpha4beta1.
Authors Bridges Lance C; Tani Patricia H; Hanson Krista R; Roberts Charles M; Judkins Matthew B; Bowditch Ron D;
JournalJ Biol Chem
PubMed ID11724793
The interaction of lymphocytes with other cells is critical for normal immune surveillance and response. MDC-L (ADAM 28), a member of the ADAM (a disintegrin and metalloprotease) protein family, is expressed on the surface of human lymphocytes. ADAMs possess a disintegrin-like domain similar in sequence to small non-enzymatic snake venom ... More
Evidence for the vectorial nature of drug (substrate)-stimulated ATP hydrolysis by human P-glycoprotein.
Authors Sauna Z E; Smith M M; Müller M; Ambudkar S V;
JournalJ Biol Chem
PubMed ID11451943
P-glycoprotein (Pgp), the ATP-binding cassette multidrug transporter, exhibits a drug (substrate)-stimulatable ATPase activity, and vanadate (Vi) inhibits this activity by stably trapping the nucleoside diphosphate in the Pgp.ADP.Vi conformation. We recently demonstrated that Vi-induced 8-azido-[alpha-(32)P]ADP trapping into Pgp in the absence of substrate occurs both in the presence of 8-azido-[alpha-(32)P]ATP ... More