'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
Identification of the major synaptojanin-binding proteins in brain.
Authorsde Heuvel E, Bell AW, Ramjaun AR, Wong K, Sossin WS, McPherson PS
JournalJ Biol Chem
PubMed ID9079704
'Synaptojanin is a nerve-terminal enriched inositol 5-phosphatase thought to function in synaptic vesicle endocytosis, in part through interactions with the Src homology 3 domain of amphiphysin. We have used synaptojanin purified from Sf9 cells after baculovirus mediated expression in overlay assays to identify two major synaptojanin-binding proteins in rat brain. ... More
Identification of a novel human eicosanoid receptor coupled to G(i/o).
'We have conducted an in silico data base search for and cloned a novel G-protein-coupled receptor (GPCR) named TG1019. Dot and Northern blotting analyses showed that transcripts of the novel GPCR were expressed in various tissues except brain, and the expression was more intense in liver, kidney, peripheral leukocyte, lung, ... More
VRILLE feeds back to control circadian transcription of Clock in the Drosophila circadian oscillator.
'The Drosophila circadian oscillator consists of interlocked period (per)/timeless (tim) and Clock (Clk) transcriptional/translational feedback loops. Within these feedback loops, CLK and CYCLE (CYC) activate per and tim transcription at the same time as they repress Clk transcription, thus controlling the opposite cycling phases of these transcripts. CLK-CYC directly bind ... More
Accurate processing and secretion in the baculovirus expression system of an erythroid-cell-stimulating factor consisting of a chimaera of insulin-like growth factor II and an insect insulin-like peptide.
AuthorsCongote LF, Li Q
JournalBiochem J
PubMed ID8166627
'A synthetic gene encoding the signal peptide and the N-terminal sequence of bombyxin, an insect insulin-like peptide, and the 58 amino acids of the C-terminal sequence of human insulin-like growth factor II (IGF II) has been expressed using the baculovirus system. This synthetic chimaera was obtained by amplification of four ... More
Superior biologic activity of the recombinant bee venom allergen hyaluronidase expressed in baculovirus-infected insect cells as compared with Escherichia coli
AuthorsSoldatova LN, Crameri R, Gmachl M, Kemeny DM, Schmidt M, Weber M, Mueller UR
JournalJ Allergy Clin Immunol
PubMed ID9600508
'BACKGROUND: Hyaluronidase (Hya) is one of several allergens in honeybee venom. Its cDNA sequence was recently described. OBJECTIVE: We sought to express recombinant Hya in prokaryotic and eukaryotic systems and to compare it with natural (n)Hya for biologic activity. METHODS: In Escherichia coli Hya was produced as inclusion body 6 ... More
Purification, reconstitution, and steady-state kinetics of the trans-membrane 17 beta-hydroxysteroid dehydrogenase 2.
Authors Lu Ming-Liang; Huang Yi-Wei; Lin Sheng-Xiang;
JournalJ Biol Chem
PubMed ID11940569
'Human membrane 17 beta-hydroxysteroid dehydrogenase 2 is an enzyme essential in the conversion of the highly active 17beta-hydroxysteroids into their inactive keto forms in a variety of tissues. 17 beta-hydroxysteroid dehydrogenase 2 with 6 consecutive histidines at its N terminus was expressed in Sf9 insect cells. This recombinant protein retained ... More
Biochemical characterization of the multifunctional Ca2+/calmodulin- dependent protein kinase type IV expressed in insect cells.
AuthorsCruzalegui FH, Means AR
JournalJ Biol Chem
PubMed ID8253736
'We have expressed the rat brain Ca2+/calmodulin (CaM)-dependent protein kinase type IV in insect cells. The recombinant enzyme is produced as a single polypeptide that migrates on SDS-polyacrylamide gel electrophoresis at 61 kDa. Recombinant CaM kinase IV undergoes slow CaM- dependent autophosphorylation. The autophosphorylation of CaM kinase IV occurs on ... More
Monitoring foreign protein expression under baculovirus p10 and polh promoters in insect larvae.
Authors Cha H J; Dalal N G; Pham M Q; Kramer S F; Vakharia V N; Bentley W E;
JournalBiotechniques
PubMed ID12019794
Monitoring foreign protein expression under baculovirus p10 and polh promoters in insect larvae.
ATP-dependent nucleosome disruption at a heat-shock promoter mediated by binding of GAGA transcription factor [see comments]
AuthorsTsukiyama T, Becker PB, Wu C
JournalNature
PubMed ID8107823
Genetic control elements are usually situated in local regions of chromatin that are hypersensitive to structural probes such as DNase I. We have reconstructed the chromatin structure of the hsp70 promoter using an in vitro nucleosome assembly system. Binding of the GAGA transcription factor on existing nucleosomes leads to nucleosome ... More
Discovery of a small molecule insulin mimetic with antidiabetic activity in mice [see comments]
AuthorsZhang B, Salituro G, Szalkowski D, Li Z, Zhang Y, Royo I, Vilella D, Diez MT, Pelaez F, Ruby C, Kendall RL, Mao X, Griffin P, Calaycay J, Zierath JR, Heck JV, Smith RG, Moller DE
JournalScience
PubMed ID10320380
Insulin elicits a spectrum of biological responses by binding to its cell surface receptor. In a screen for small molecules that activate the human insulin receptor tyrosine kinase, a nonpeptidyl fungal metabolite (L-783,281) was identified that acted as an insulin mimetic in several biochemical and cellular assays. The compound was ... More
A lymphotoxin-beta-specific receptor [see comments]
AuthorsCrowe PD, VanArsdale TL, Walter BN, Ware CF, Hession C, Ehrenfels B, Browning JL, Din WS, Goodwin RG, Smith CA
JournalScience
PubMed ID8171323
Tumor necrosis factor (TNF) and lymphotoxin-alpha (LT-alpha) are members of a family of secreted and cell surface cytokines that participate in the regulation of immune and inflammatory responses. The cell surface form of LT-alpha is assembled during biosynthesis as a heteromeric complex with lymphotoxin-beta (LT-beta), a type II transmembrane protein ... More
The ubiquitous chromatin protein DEK alters the structure of DNA by introducing positive supercoils.
We have investigated the molecular mechanism by which the proto-oncogene protein DEK, an abundant chromatin-associated protein, changes the topology of DNA in chromatin in vitro. Band-shift assays and electron microscopy revealed that DEK induces both intra- and intermolecular interactions between DNA molecules. Binding of the DEK protein introduces constrained positive ... More
Genetic screens in yeast to identify mammalian nonreceptor modulators of G-protein signaling.
AuthorsCismowski MJ, Takesono A, Ma C, Lizano JS, Xie X, Fuernkranz H, Lanier SM, Duzic E
JournalNature Biotechnology
PubMed ID10471929
We describe genetic screens in Saccharomyces cerevisiae designed to identify mammalian nonreceptor modulators of G-protein signaling pathways. Strains lacking a pheromone-responsive G-protein coupled receptor and expressing a mammalian-yeast Galpha hybrid protein were made conditional for growth upon either pheromone pathway activation (activator screen) or pheromone pathway inactivation (inhibitor screen). Mammalian ... More
Use of the baculovirus system to assemble polyomavirus capsid-like particles with different polyomavirus structural proteins: analysis of the recombinant assembled capsid-like particles.
AuthorsAn K, Gillock ET, Sweat JA, Reeves WM, Consigli RA
JournalJ Gen Virol
PubMed ID10211971
The genes encoding the structural proteins (VP1, VP2 and VP3) of murine polyomavirus were cloned into the p2Bac dual multiple cloning site vector, individually or jointly, and the corresponding proteins were expressed in Spodoptera frugiperda (Sf9) insect cells by cotransfecting Sf9 cells with the constructed vector and the linear DNA ... More
Recombinant murine serum amyloid A from baculovirus-infected insect cells: purification and characterization
AuthorsKluve-Beckerman B, Song M, Benson MD, Liepnieks JJ
JournalBiochim Biophys Acta
PubMed ID8399365
Serum amyloid A (SAA) is an extremely sensitive acute-phase reactant and precursor to the subunit protein in reactive amyloid deposits. Although the mouse has long served as an informative experimental model, both the function of SAA and the pathogenic mechanism of amyloid formation remain unknown. The production of SAA by ... More
Enzymatic properties of human Na,K-ATPase alpha1beta3 isozyme.
AuthorsYu C, Xie Z, Askari A, Modyanov NN
JournalArch Biochem Biophys
PubMed ID9281321
Recent results of a wide-scale human cDNA sequencing project have identified a cDNA which encodes a hitherto unknown human protein sequence exhibiting structural similarities with beta-subunits of the Na,K- and H,K-ATPase family and with the amphibian Na,KATPase beta3-subunit, in particular. In this study the ability of the putative human beta3-subunit ... More
Polyomavirus major capsid protein VP1 is capable of packaging cellular DNA when expressed in the baculovirus system
AuthorsGillock ET, Rottinghaus S, Chang D, Cai X, Smiley SA, An K, Consigli RA
JournalJ Virol
PubMed ID9060642
Using the p2Bac dual multiple cloning site transfer vector, the polyomavirus major capsid protein gene VP1 was cloned for expression in the baculovirus-insect cell expression system. The 5-day-infected cellular lysate from this recombinant preparation was purified by cesium chloride density gradient centrifugation. Capsid-like particles were observed in the resulting preparation. ... More
Regulation of Vav-SLP-76 binding by ZAP-70 and its relevance to TCR zeta/CD3 induction of interleukin-2.
AuthorsRaab M, da Silva AJ, Findell PR, Rudd CE
JournalImmunity
PubMed ID9047237
T cell activation stimulates p56(lck), p59(fyn), ZAP-70, Vav-SLP-76 binding, and IL-2 transcription. Major questions concern the tyrosine- kinase and relevant site(s) needed for Vav-SLP-76 complex formation and its role in IL-2 production. Here, we show that of the three kinases, only ZAP-70 phosphorylates SLP-76 at specific sites that allow Vav ... More
The Bmx tyrosine kinase induces activation of the Stat signaling pathway, which is specifically inhibited by protein kinase Cdelta.
AuthorsSaharinen P, Ekman N, Sarvas K, Parker P, Alitalo K, Silvennoinen O
JournalBlood
PubMed ID9373245
Members of the hematopoietically expressed Tec tyrosine kinase family have an important role in hematopoietic signal transduction, as exemplified by the crucial role of Btk for B-cell differentiation and activation. Although a variety of cell surface receptors have been found to activate Tec tyrosine kinases, the specific signaling pathways and ... More
Direct expression of the pig testicular 3 alpha/beta(20 beta)- hydroxysteroid dehydrogenase cDNA using baculovirus expression system
AuthorsNakajin S, Nakajima T, Uchida M, Shinoda M
JournalYakugaku Zasshi
PubMed ID7602499
The direct expression of the pig testicular 3 alpha/beta(20 beta)- hydroxysteroid dehydrogenase (HSD) cDNA using a baculovirus expression system was investigated. A minimum essential cDNA containing the coding region of 3 alpha/beta(20 beta)-HSD was amplified by polymerase chain reaction (PCR) to extend the DNA linker including the Bam HI restriction ... More
Functional characterization of partially purified Epstein-Barr virus DNA polymerase expressed in the baculovirus system.
AuthorsLin JC, De BK, Mar EC
JournalVirus Genes
PubMed ID7975269
The DNA polymerase gene of Epstein-Barr virus (EBV) was cloned into baculovirus transfer vector (pBlueBac). The recombinant baculovirus (AcEBP-15) was obtained by cotransfection of Spodoptera frugiperda (Sf9) cells with infectious DNA from Autographa californica multiple nuclear polyhedrin virus (AcMNPV) and pBlueBac plasmid carrying EBV polymerase gene. Infection of Sf9 cells ... More
Expression, purification and characterization of recombinant C. Elegans UNC-18.
AuthorsOgawa H, Hayashi N, Hori I, Kobayashi T, Hosono R
JournalNeurochem Int
PubMed ID8939465
The Caenorhabditis elegans unc-18-encoded protein (UNC-18) is implicated in the processes of vesicle targeting, docking, and/or fusion. To further characterize the properties of this important neural protein, we expressed it at a high level in Spodoptera frugiperda Sf21 cells using a baculovirus expressing system. A cDNA containing the coding sequence ... More
Interactions of CBL with BCR-ABL and CRKL in BCR-ABL-transformed myeloid cells.
AuthorsBhat A, Kolibaba K, Oda T, Ohno-Jones S, Heaney C, Druker BJ
JournalJ Biol Chem
PubMed ID9195915
The Philadelphia chromosome, detected in virtually all cases of chronic myelogenous leukemia (CML), is formed by a reciprocal translocation between chromosomes 9 and 22 that fuses BCR-encoded sequences upstream of exon 2 of c-ABL. The BCR-ABL fusion creates a gene whose protein product, p210BCR-ABL, has been implicated as the cause ... More
Determination of substrate motifs for human Chk1 and hCds1/Chk2 by the oriented peptide library approach.
Authors O'Neill Ted; Giarratani Lauren; Chen Ping; Iyer Lakshmanan; Lee Chang-Hun; Bobiak Matthew; Kanai Fumihiko; Zhou Bin-Bing; Chung Jay H; Rathbun Gary A;
JournalJ Biol Chem
PubMed ID11821419
Mammalian Chk1 and Chk2 are two Ser/Thr effector kinases that play critical roles in DNA damage-activated cell cycle checkpoint signaling pathways downstream of ataxia telangiectasia-mutated and ataxia telangiectasia-related. Endogenous substrates have been identified for human hCds1/Chk2 and Chk1; however, the sequences surrounding the substrate residues appear unrelated, and consensus substrate ... More
Mutations Targeted to a Predicted Helix in the Extreme Carboxyl-terminal Region of the Human Estrogen Receptor-alpha Alter Its Response to Estradiol and 4-Hydroxytamoxifen.
The human estrogen receptor-alpha, a member of the nuclear receptor superfamily, is a ligand-regulated transcriptional modulator. Because comparatively little is known about the extreme carboxyl-terminal region of the estrogen receptor (F domain), we used secondary structure prediction to design mutations that delete the F domain (S554stop), disrupt a possible turn ... More