Process formation in Sf9 cells induced by the expression of a microtubule-associated protein 2C-like construct.
AuthorsLeClerc N, Kosik KS, Cowan N, Pienkowski TP, Baas PW
JournalProc Natl Acad Sci U S A
PubMed ID8327502
'To understand the roles of various microtubule-associated proteins (MAPs) in the development of axons and dendrites, we have expressed individual neuronal MAPs in normally rounded Sf9 host cells. We previously reported that expression of tau protein in these cells results in the elaboration of long processes containing dense bundles of ... More
Insulin receptor substrate 1 mediates insulin and insulin-like growth factor I-stimulated maturation of Xenopus oocytes.
'Insulin and insulin-like growth factor I (IGF-I) initiate cellular functions by activating their homologous tyrosine kinase receptors. In most mammalian cell types, this results in rapid tyrosine phosphorylation of a high-molecular-weight substrate termed insulin receptor substrate 1 (IRS-1). Previous studies suggest that IRS-1 may act as a docking protein that ... More
Molecular cloning and characterization of Porphyromonas gingivalis lysine-specific gingipain. A new member of an emerging family of pathogenic bacterial cysteine proteinases.
'The proteinases of Porphyromonas gingivalis are key virulence factors in the etiology and progression of periodontal disease. Previous work in our laboratories resulted in the purification of arginine- and lysine-specific cysteine proteinases, designated gingipains, that consist of several tightly associated protein subunits. Recent characterization of arginine-specific gingipain-1 (gingipain R1; RGP-1) ... 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
Endotoxin-mediated endothelial cell injury and activation: role of soluble CD14.
AuthorsArditi M, Zhou J, Dorio R, Rong GW, Goyert SM, Kim KS
JournalInfect Immun
PubMed ID7687581
'Vascular endothelial cell (EC) injury by lipopolysaccharides (LPS) plays a major role in the pathogenesis of gram-negative bacterial sepsis and endotoxic shock. The studies described here were performed to define further the molecular mechanisms involved in the EC responses to LPS. We showed that serum was required for LPS-mediated cytotoxicity ... 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
Production of a recombinant imported fire ant venom allergen, Sol i 2, in native and immunoreactive form.
AuthorsSchmidt M, McConnell TJ, Hoffman DR
JournalJ Allergy Clin Immunol
PubMed ID8765821
'BACKGROUND: The complementary DNA encoding for the important imported fire ant venom allergen, Sol i 2, has previously been cloned. The binding of human IgE antibodies to Sol i 2 has been demonstrated to be conformation-dependent. METHODS: A couple cDNA clone encoding the Sol i 2 protein sequence and its ... 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
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
Calcium-mediated Association of a Putative Vacuolar Sorting Receptor PV72 with a Propeptide of 2S Albumin.
'PV72, a type I membrane protein with three epidermal-growth factor (EGF)-like motifs, was found to be localized on the membranes of the precursor-accumulating (PAC) vesicles that accumulated precursors of various seed storage proteins. To clarify the function of PV72 as a sorting receptor, we expressed four modified PV72s and analyzed ... More
Selective reconstitution and recovery of functional gamma-secretase complex on budded baculovirus particles.
AuthorsHayashi I, Urano Y, Fukuda R, Isoo N, Kodama T, Hamakubo T, Tomita T, Iwatsubo T,
JournalJ Biol Chem
PubMed ID15215237
'In vitro reconstitution of functions of membrane proteins is often hampered by aggregation, misfolding, or lack of post-translational modifications of the proteins attributable to overexpression. To overcome this technical obstacle, we have developed a method to express multimeric integral membrane proteins in extracellular (budded) baculovirus particles that are released from ... 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
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
Identification of the ligand-binding regions in the macrophage colony- stimulating factor receptor extracellular domain.
AuthorsWang ZE, Myles GM, Brandt CS, Lioubin MN, Rohrschneider L
JournalMol Cell Biol
PubMed ID8355686
The c-fms gene encodes the receptor for the macrophage colony- stimulating factor (M-CSF), and its extracellular domain consists of five immunoglobulin-like subdomains. To identify which of the five immunoglobulin-like regions are involved in ligand binding, we polymerase chain reaction-cloned five segments of the extracellular domain of the murine c-fms gene, ... More
Expression of bovine vitamin K-dependent carboxylase activity in baculovirus-infected insect cells.
AuthorsRoth DA, Rehemtulla A, Kaufman RJ, Walsh CT, Furie B, Furie BC
JournalProc Natl Acad Sci U S A
PubMed ID8378308
A vitamin K-dependent carboxylase has recently been purified from bovine liver microsomes and candidate cDNA clones have been isolated. Definitive identification of the carboxylase remains circumstantial since expression of candidate carboxylase cDNAs in mammalian cells is confounded by the presence of endogenous carboxylase activity. To overcome this problem, a recombinant ... More
Human IL-1 beta processing and secretion in recombinant baculovirus- infected Sf9 cells is blocked by the cowpox virus serpin crmA.
Biologically active, mature IL-1 beta (mIL-1 beta) is released from activated monocytes after proteolytic processing from an inactive precursor (pIL-1 beta). IL-1 beta converting enzyme (ICE), the first member of a newly discovered family of cysteine proteinases, is required for this processing event. The cleaved cytokine is released from monocytes ... 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
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
Heparin-binding protein (CAP37) is internalized in monocytes and increases LPS-induced monocyte activation.
AuthorsHeinzelmann M, Mercer-Jones MA, Flodgaard H, Miller FN
JournalJ Immunol
PubMed ID9605157
Previous studies have shown that the neutrophil-derived heparin-binding protein (HBP), also known as CAP37 or azurocidin, potentiates the LPS- induced release of proinflammatory cytokines (TNF-alpha, IL-1, and IL- 6) from isolated human monocytes. To date, the mechanisms by which HBP enhances LPS-induced monocyte activation have not been elucidated, and it ... More
Purification and characterization of the human elongator complex.
Authors Hawkes Nicola A; Otero Gabriel; Winkler G Sebastiaan; Marshall Nick; Dahmus Michael E; Krappmann Daniel; Scheidereit Claus; Thomas Claire L; Schiavo Giampietro; Erdjument-Bromage Hediye; Tempst Paul; Svejstrup Jesper Q;
JournalJ Biol Chem
PubMed ID11714725
Human Elongator complex was purified to virtual homogeneity from HeLa cell extracts. The purified factor can exist in two forms: a six-subunit complex, holo-Elongator, which has histone acetyltransferase activity directed against histone H3 and H4, and a three-subunit core form, which does not have histone acetyltransferase activity despite containing the ... 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