Mechanism and structure based inhibitors of phospholipase C enzymes.
AuthorsRoberts MF, Wu Y, Zhou C, Geng D, Tan C
JournalAdv Enzyme Regul
PubMed ID8869740
'PI-specific PLC enzymes are a key component of phosphatidylinositol-mediated signaling pathways since the hydrophobic product, diacylglycerol, activates protein kinase C and the water-soluble product, inositol trisphosphate, is involved in Ca2+ mobilization. Nonspecific, or PC-PLC, enzymes can generate diacylglycerol without Ca2+ mobilization. A series of inhibitors, both lipophilic and water-soluble, have ... More
Diacylglycerol induces fusion of nuclear envelope membrane precursor vesicles.
'Purified membrane vesicles isolated from sea urchin eggs form nuclear envelopes around sperm nuclei following GTP hydrolysis in the presence of cytosol. A low density subfraction of these vesicles (MV1), highly enriched in phosphatidylinositol (PtdIns), is required for nuclear envelope formation. Membrane fusion of MV1 with a second fraction that ... More
A turbidometric assay for phospholipase C and sphingomyelinase.
AuthorsTorley L, Silverstrim C, Pickett W
JournalAnal Biochem
PubMed ID7864373
'We describe a simple turbidometric assay for phosphatidylcholine-specific phospholipase C (PC-PLC) (EC 3.1.4.3), phosphatidylinositol-specific phospholipase C (PI-PLC) (EC 3.1.4.10), and sphingomyelinase (SMase) (EC 3.1.4.12), suitable for high-volume screening using unmodified substrates. Under the conditions described, 1 to 10 U/ml of PC-PLC (Bacillus cereus) induces a rapid and continuous increase in ... More
Design, synthesis, and evaluation of water-soluble phospholipid analogues as inhibitors of phospholipase C from Bacillus cereus.
AuthorsFranklin CL, Li H, Martin SF
JournalJ Org Chem
PubMed ID12968879
'The rate of hydrolysis of natural phospholipids by the phosphatidylcholine-preferring phospholipase C from Bacillus cereus (PLC(Bc)) follows the order phosphatidylcholine > phosphatidylethanolamine >> phosphatidyl-l-serine. To probe the structural basis for this substrate specificity, a series of water-soluble, nonhydrolyzable substrate analogues were needed so their complexes with the enzyme could be ... More
Internalization of exogenously added memapsin 2 (beta-secretase) ectodomain by cells is mediated by amyloid precursor protein.
'Memapsin 2 (beta-secretase) is the protease that initiates cleavage of amyloid precursor protein (APP) leading to the production of amyloid-beta (Abeta) peptide and the onset of Alzheimer''s disease. Both APP and memapsin 2 are Type I transmembrane proteins and are endocytosed into endosomes where APP is cleaved by memapsin 2. ... More
A high level of cell surface phosphatidylinositol-specific phospholipase C activity is characteristic of growth-arrested 3T3 fibroblasts but not of transformed variants.
'Confluent monolayers of four contact-inhibited mouse fibroblast lines (Swiss 3T3, Balb/c 3T3, NIH 3T3, and C3H10T1/2) were found to have substantial levels of a cell surface phosphatidylinositol-specific phospholipase C (ecto-PLC). In contrast, confluent cultures of virally, chemically, or spontaneously transformed variants derived from these cell lines expressed undetectable or negligible ... More
Crystal structure of the phosphatidylinositol-specific phospholipase C from Bacillus cereus in complex with myo-inositol.
AuthorsHeinz DW, Ryan M, Bullock TL, Griffith OH
JournalEMBO J
PubMed ID7664726
'Phosphatidylinositol (PI), once regarded as an obscure component of membranes, is now recognized as an important reservoir of second messenger precursors and as an anchor for membrane enzymes. PI-specific phospholipase C (PI-PLC) is the enzyme that cleaves PI, invoking numerous cellular responses. The crystal structure of PI-PLC from Bacillus cereus ... More
Kinetics of phosphatidylinositol-specific phospholipase C with vesicles of a thiophosphate analogue of phosphatidylinositol.
AuthorsHendrickson HS, Banovetz C, Kirsch MJ, Hendrickson EK
JournalChem Phys Lipids
PubMed ID9022217
'1,2-Dimyristoyloxypropane-3-thiophospho(1D-1-myo-inositol) (D-thio-DMPI) was synthesized as a substrate for the continuous spectrophotometric assay of phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus cereus. Release of thio-diglyceride is followed by a coupled reaction with 4,4''-dithiopyridine to produce a chromophore, 4-thiopyridine, measured by its absorption at 324 nm. Sonicated vesicles of D-thio-DMPI gave sigmoidal Michaelis-Menten ... More
A surface antigen of Giardia lamblia with a glycosylphosphatidylinositol anchor.
AuthorsDas S, Traynor-Kaplan A, Reiner DS, Meng TC, Gillin FD
JournalJ Biol Chem
PubMed ID1657957
'Since Giardia lamblia trophozoites are exposed to high concentrations of fatty acids in their human small intestinal milieu, we determined the pattern of incorporation of [3H]palmitic acid and myristic acid into G. lamblia proteins. The pattern of fatty acylation was unusually simple since greater than 90% of the Giardia protein ... More
Phospholipids chiral at phosphorus. Stereochemical mechanism of reactions catalyzed by phosphatidylinositide-specific phospholipase C from Bacillus cereus and guinea pig uterus.
AuthorsLin GL, Bennett CF, Tsai MD
JournalBiochemistry
PubMed ID2161255
'(Rp)- and (Sp)-1,2-dipalmitoyl-sn-glycero-3-thiophosphoinositol (DPPsI) were synthesized as a mixture and their configurations assigned on the basis of the stereospecific hydrolysis catalyzed by phospholipase A2 (PLA2) from bee venom. PLA2 is known to be stereospecific to the Rp isomer of 1,2-dipalmitoyl-sn-glycero-3-thiophosphocholine (DPPsC) and 1,2-dipalmitoyl-sn-glycero-3-thiophosphoethanolamine (DPPsE). Since the configurations of (Rp)- and ... More
Glycosyl-phosphatidylinositol anchored acetylcholinesterase as substrate for phosphatidylinositol-specific phospholipase C from Bacillus cereus.
AuthorsStieger S, Brodbeck U
JournalBiochimie
PubMed ID1660725
'We investigated the enzymatic properties of phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus cereus towards glycosyl-phosphatidylinositol anchored acetylcholinesterase (AChE) from bovine erythrocytes and Torpedo electric organ as substrate. The conversion of membrane from AChE to soluble AChE by PI-PLC depended on the presence of a detergent and of phosphatidylcholine. In presence ... More
Interactions between saturated acyl chains confer detergent resistance on lipids and glycosylphosphatidylinositol (GPI)-anchored proteins: GPI-anchored proteins in liposomes and cells show similar behavior.
AuthorsSchroeder R, London E, Brown D
JournalProc Natl Acad Sci U S A
PubMed ID7991596
'Proteins anchored by GPI are poorly solubilized from cell membranes by cold nonionic detergents because they associate with detergent-resistant membranes rich in cholesterol and sphingolipids. In this study, we demonstrated that cholesterol and sphingolipid-rich liposomes were incompletely solubilized by Triton X-100. GPI-anchored placental alkaline phosphatase incorporated in these liposomes was ... More
Phase partitioning detects differences between phospholipase-released forms of alkaline phosphatase--a GPI-linked protein.
AuthorsRaymond FD, Moss DW, Fisher D
JournalBiochim Biophys Acta
PubMed ID7678987
'A number of enzymes are known to release alkaline phosphatase and other glycan phosphatidylinositol-anchored proteins from membrane surfaces. We describe a novel approach to detect and measure these activities by partitioning in aqueous phase systems. The procedures avoid the complications of micelle-formation involving hydrophobic molecules that may arise with detergent-based ... More
Phosphatidylinositol-specific phospholipases C from Bacillus cereus and Bacillus thuringiensis.
AuthorsGriffith OH, Volwerk JJ, Kuppe A
JournalMethods Enzymol
PubMed ID1646937
Toward the mechanism of phosphoinositide-specific phospholipases C.
AuthorsBruzik KS, Tsai MD
JournalBioorg Med Chem
PubMed ID7922124
Functional characteristics of phosphatidylinositol-specific phospholipases C from Bacillus cereus and Bacillus thuringiensis.
Phosphatidylinositol-specific phospholipase C was purified from the culture medium of B. thuringiensis to high specific activity using a procedure we recently described for purification of PI-PLC from B. cereus (Volwerk et al. (1989) J. Cell. Biochem. 39, 315-325). The purified enzymes from B. thuringiensis and B. cereus have similar specific ... More
Renal dipeptidase is one of the membrane proteins released by phosphatidylinositol-specific phospholipase C.
AuthorsHooper NM, Low MG, Turner AJ
JournalBiochem J
PubMed ID2822007
Renal dipeptidase (dehydropeptidase-I, EC 3.4.13.11) was released from pig kidney membrane preparations by treatment with phosphatidylinositol-specific phospholipase C from Staphylococcus aureus and Bacillus thuringiensis and a phospholipase C preparation from Bacillus cereus to a similar extent as alkaline phosphatase. Endopeptidase-24.11 and aminopeptidase N were not released by this treatment. After ... More
Activity of phosphatidylinositol-specific phospholipase C from Bacillus cereus with thiophosphate analogs of dimyristoylphosphatidylinositol.
AuthorsHendrickson HS, Giles AN, Vos SE
JournalChem Phys Lipids
PubMed ID9353901
Phosphatidylinositol-specific phospholipase C (PI-PLC) was studied with sonicated dispersions of a thiophosphate analog of phosphatidylinositol, 1, 2-dimyristoyloxypropane-3-thiophospho(1D-1-myo-inositol) (D-thio-DMPI). Kinetic parameters were derived from the rate as a function of bulk lipid concentration at constant saturating surface concentration of substrate (case I), and as a function of surface concentration of substrate ... More
Crystallization of phosphatidylinositol-specific phospholipase C from Bacillus cereus.
AuthorsBullock TL, Ryan M, Kim SL, Remington SJ, Griffith OH
JournalBiophys J
PubMed ID8386017
Phosphatidylinositol-specific phospholipase C (PI-PLC) cleaves phosphoinositides into two parts, lipid-soluble diacylglycerol and the water-soluble phosphorylated inositol. Two crystal forms of Bacillus cereus PI-PLC have been obtained by the vapor diffusion technique. Hexagonal crystals were grown from solutions containing polyethylene glycol (PEG; 4,000 to 8,000 D). The space group of these ... More
Mammalian cells that express Bacillus cereus phosphatidylinositol-specific phospholipase C have increased levels of inositol cyclic 1:2-phosphate, inositol 1-phosphate, and inositol 2-phosphate.
AuthorsRoss TS, Wang FP, Majerus PW
JournalJ Biol Chem
PubMed ID1328186
Phosphatidylinositol-specific phospholipase C (PtdIns-PLC) of Bacillus cereus catalyzes the conversion of PtdIns to inositol cyclic 1:2-phosphate and diacylglycerol. NIH 3T3, Swiss mouse 3T3, CV-1, and Cos-7 cells were transfected with a cDNA encoding this enzyme, and the metabolic and cellular consequences were investigated. Overexpression of PtdIns-PLC enzyme activity was associated ... More
Mutagenesis study of the glycosylphosphatidylinositol phospholipase C of Trypanosoma brucei.
AuthorsCarnall N, Webb H, Carrington M
JournalMol Biochem Parasitol
PubMed ID9476790
The glycosylphosphatidylinositol phospholipase C (GPI-PLC) from Trypanosoma brucei is particularly effective in hydrolysing the GPI-anchors of some proteins. The enzyme is inhibited by Zn2+ and p-chloromercurylphenylsulphonic acid, both of which can act as sulphydryl reagents, suggesting that a cysteine residue may be important in catalysis. Single cysteine to serine mutants ... More
Substrate stereospecificity of phosphatidylinositol-specific phospholipase C from Bacillus cereus examined using the resolved enantiomers of synthetic myo-inositol 1-(4-nitrophenyl phosphate).
The substrate stereospecificity of phosphatidylinositol-specific phospholipase C from Bacillus cereus is examined using the resolved optical isomers of synthetic myo-inositol 1-(4-nitrophenyl phosphate), a chromogenic substrate for the phospholipase. The synthetic route employs mild acid-labile protecting groups and separation of the substituted myo-inositol enantiomers as the (-)-camphanyl ester diastereomers. Measurements of ... More
Allosteric interactions within subsites of a monomeric enzyme: kinetics of fluorogenic substrates of PI-specific phospholipase C.
AuthorsBirrell GB, Zaikova TO, Rukavishnikov AV, Keana JF, Griffith OH
JournalBiophys J
PubMed ID12719256
Two novel water-soluble fluorescein myo-inositol phosphate (FLIP) substrates, butyl-FLIP and methyl-FLIP, were used to examine the kinetics and subsite interactions of Bacillus cereus phosphatidylinositol-specific phospholipase C. Butyl-FLIP exhibited sigmoidal kinetics when initial rates are plotted versus substrate concentration. The data fit a Hill coefficient of 1.2-1.5, suggesting an allosteric interaction ... More
Removal or maintenance of inositol-linked acyl chain in glycosylphosphatidylinositol is critical in trypanosome life cycle.
AuthorsHong Y, Nagamune K, Morita YS, Nakatani F, Ashida H, Maeda Y, Kinoshita T
JournalJ Biol Chem
PubMed ID16510441
The protozoan parasite Trypanosoma brucei is coated by glycosylphosphatidylinositol (GPI)-anchored proteins. During GPI biosynthesis, inositol in phosphatidylinositol becomes acylated. Inositol is deacylated prior to attachment to variant surface glycoproteins in the bloodstream form, whereas it remains acylated in procyclins in the procyclic form. We have cloned a T. brucei GPI ... More
Human erythrocyte sialidase is linked to the plasma membrane by a glycosylphosphatidylinositol anchor and partly located on the outer surface.
AuthorsChiarini A, Fiorilli A, Di Francesco L, Venerando B, Tettamanti G
JournalGlycoconj J
PubMed ID8395269
Treatment of human erythrocyte ghosts with phosphatidylinositol-phospholipase C (PIPLC) from Bacillus cereus liberated the ghost-linked sialidase. Maximal release of sialidase (about 70% of total) was achieved by incubating ghosts at 37 degrees C for 60 min, at pH 6.0, with PIPLC (PIPLC total units/ghost protein ratio, 4.5 each time) added ... More
Phosphatidylinositol-specific phospholipase C of Bacillus cereus: cloning, sequencing, and relationship to other phospholipases.
AuthorsKuppe A, Evans LM, McMillen DA, Griffith OH
JournalJ Bacteriol
PubMed ID2509427
The phosphatidylinositol (PI)-specific phospholipase C (PLC) of Bacillus cereus was cloned into Escherichia coli by using monoclonal antibody probes raised against the purified protein. The enzyme is specific for hydrolysis of the membrane lipid PI and PI-glycan-containing membrane anchors, which are important structural components of one class of membrane proteins. ... More
Effects of intra- and extracellularly applied phospholipases C on excitability of squid giant axons.
AuthorsNakajima M, Ikezawa H, Yamagishi S
JournalToxicon
PubMed ID2877512
The effects of phospholipases C on the membrane excitability of the squid giant axon were investigated using phosphatidylcholine-hydrolyzing phospholipase C and sphingomyelinase C of Bacillus cereus, and phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis. When the squid axon was perfused internally with phosphatidylcholine-hydrolyzing phospholipase C in KF or K-glutamate solution, the ... More
Cloning, expression, and mutagenesis of phosphatidylinositol-specific phospholipase C from Staphylococcus aureus: a potential staphylococcal virulence factor.
AuthorsDaugherty S, Low MG
JournalInfect Immun
PubMed ID8225585
Staphylococcus aureus secretes a phosphatidylinositol (PI)-specific phospholipase C (PI-PLC) which is able to hydrolyze the membrane lipid PI and membrane protein anchors containing glycosyl-PI. The gene for PI-PLC (plc) was cloned from S. aureus into Escherichia coli. Oligonucleotide probes based on partial protein sequence and polyclonal antibodies raised against the ... More
HDL3 binds to glycosylphosphatidylinositol-anchored proteins to activate signalling pathways.
Previous studies have indicated that in HepG2 cells HDL3-signalling involves glycosylphosphatidylinositol (GPI) anchored proteins. HDL3-binding to HepG2 cells was found to be enhanced by cellular preincubation with PI-PLC inhibitors and sensitive to a cellular preincubation with exogenous PI-PLC, suggesting that HDL3 binds directly on GPI-anchored proteins to initiate signaling. Moreover ... More
Determination of pKa values of the histidine side chains of phosphatidylinositol-specific phospholipase C from Bacillus cereus by NMR spectroscopy and site-directed mutagenesis.
AuthorsLiu T, Ryan M, Dahlquist FW, Griffith OH
JournalProtein Sci
PubMed ID9300493
Two active site histidine residues have been implicated in the catalysis of phosphatidylinositol-specific phospholipase C (PI-PLC). In this report, we present the first study of the pKa values of histidines of a PI-PLC. All six histidines of Bacillus cereus PI-PLC were studied by 2D NMR spectroscopy and site-directed mutagenesis. The ... More
Probing the roles of active site residues in phosphatidylinositol-specific phospholipase C from Bacillus cereus by site-directed mutagenesis.
AuthorsGässler CS, Ryan M, Liu T, Griffith OH, Heinz DW
JournalBiochemistry
PubMed ID9335537
The role of amino acid residues located in the active site pocket of phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus cereus[Heinz, D. W., Ryan, M., Bullock, T., & Griffith, O. H. (1995) EMBO J. 14, 3855-3863] was investigated by site-directed mutagenesis, kinetics, and crystal structure analysis. Twelve residues involved in catalysis ... More
Inhibition of the phosphatidylinositol-specific phospholipase C from Bacillus cereus by a monoclonal antibody binding to a region with sequence similarity to eukaryotic phospholipases.
AuthorsKuppe A, Hedberg KK, Volwerk JJ, Griffith OH
JournalBiochim Biophys Acta
PubMed ID1701099
Bacterial phosphatidylinositol-specific phospholipases C (PI-PLC) display similar substrate specificity as their eukaryotic counterparts involved in signal transduction of insulin and Ca2(+)-mobilizing hormones, and are used in the study of the novel glycosylphosphatidylinositol-protein anchors (GPI-anchors). For the investigation of structure-function aspects of the PI-PLC secreted from Bacillus cereus cells, a panel ... More
Modulation of PI-specific phospholipase C by membrane curvature and molecular order.
AuthorsAhyayauch H, Villar AV, Alonso A, Goñi FM
JournalBiochemistry
PubMed ID16114896
Phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus cereus has been assayed on large and small unilamellar vesicles consisting of PI, either pure or in mixtures with other lipids. Vesicle diameter (in the 50-300 nm range) influences PI-PLC activity, enzyme rates increasing with decreasing curvature radii. With sonicated unilamellar vesicles of pure ... More
Structural and mechanistic comparison of prokaryotic and eukaryotic phosphoinositide-specific phospholipases C.
AuthorsHeinz DW, Essen LO, Williams RL
JournalJ Mol Biol
PubMed ID9466937
Phosphoinositide-specific phospholipases C (PI-PLCs) are ubiquitous enzymes that catalyse the hydrolysis of phosphoinositides to inositol phosphates and diacylglycerol (DAG). Whereas the eukaryotic PI-PLCs play a central role in most signal transduction cascades by producing two second messengers, inositol-1,4,5-trisphosphate and DAG, prokaryotic PI-PLCs are of interest because they act as virulence ... More
Synthesis, structure-activity relationships, and the effect of polyethylene glycol on inhibitors of phosphatidylinositol-specific phospholipase C from Bacillus cereus.
AuthorsRyan M, Smith MP, Vinod TK, Lau WL, Keana JF, Griffith OH
JournalJ Med Chem
PubMed ID8893831
Substrate analog inhibitors of Bacillus cereus phosphatidylinositol-specific phospholipase C (PI-PLC) were synthesized and screened for their suitability to map the active site region of the enzyme by protein crystallography. Analogs of the natural substrate phosphatidylinositol (PI) were designed to examine the importance of the lipid portion and the inositol phosphate ... More
Phosphatidylinositol-specific phospholipase C from Bacillus cereus at the lipid-water interface: interfacial binding, catalysis, and activation.
AuthorsVolwerk JJ, Filthuth E, Griffith OH, Jain MK
JournalBiochemistry
PubMed ID8142343
Binding characteristics of phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus cereus binding to the phospholipid-water interface were determined by spectroscopic methods and correlated with PI-PLC's catalytic properties. Binding of the enzyme to micelles and bilayers of zwitterionic phosphocholines is accompanied by an increase in the fluorescence emission from tryptophan, whereas a ... More
The solubilization of tetrameric alkaline phosphatase from human liver and its conversion into various forms by phosphatidylinositol phospholipase C or proteolysis.
AuthorsHawrylak K, Stinson RA
JournalJ Biol Chem
PubMed ID2844768
When membrane-bound human liver alkaline phosphatase was treated with a phosphatidylinositol (PI) phospholipase C obtained from Bacillus cereus, or with the proteases ficin and bromelain, the enzyme released was dimeric. Butanol extraction of the plasma membranes at pH 7.6 yielded a water-soluble, aggregated form that PI phospholipase C could also ... More
Phospholipids chiral at phosphorus. Stereochemical mechanism for the formation of inositol 1-phosphate catalyzed by phosphatidylinositol-specific phospholipase C.
The phosphatidylinositol-specific phospholipase C (PI-PLC) from mammalian sources catalyzes the simultaneous formation of both inositol 1,2-cyclic phosphate (IcP) and inositol 1-phosphate (IP). It has not been established whether the two products are formed in sequential or parallel reactions, even though the latter has been favored in previous reports. This problem ... More
High-level expression in Escherichia coli and rapid purification of phosphatidylinositol-specific phospholipase C from Bacillus cereus and Bacillus thuringiensis.
AuthorsKoke JA, Yang M, Henner DJ, Volwerk JJ, Griffith OH
JournalProtein Expr Purif
PubMed ID1668269
The construction of four vectors for high-level expression in Escherichia coli of the phosphatidylinositol-specific phospholipase C from Bacillus cereus or Bacillus thuringiensis is described. In all constructs the coding sequence for the mature phospholipase is precisely fused to the E. coli heat-stable enterotoxin II signal sequence for targeting of the ... More
Inhibition of phosphatidylinositol-specific phospholipase C: studies on synthetic substrates, inhibitors and a synthetic enzyme.
AuthorsVizitiu D, Kriste AG, Campbell AS, Thatcher GR
JournalJ Mol Recognit
PubMed ID8877813
Enzyme inhibition studies on phosphatidylinositol-specific phospholipase C (PI-PLC) from B. Cereus were performed in order to gain an understanding of the mechanism of the PI-PLC family of enzymes and to aid inhibitor design. Inhibition studies on two synthetic cyclic phosphonate analogues (1,2) of inositol cyclic-1:2-monophosphate (cIP), glycerol-2-phosphate and vanadate were ... More
Crystal structure of phosphatidylinositol-specific phospholipase C from Bacillus cereus in complex with glucosaminyl(alpha 1-->6)-D-myo-inositol, an essential fragment of GPI anchors.
AuthorsHeinz DW, Ryan M, Smith MP, Weaver LH, Keana JF, Griffith OH
JournalBiochemistry
PubMed ID8755729
Numerous proteins on the external surface of the plasma membrane are anchored by glycosylated derivatives of phosphatidylinositol (GPI), rather than by hydrophobic amino acids embedded in the phospholipid bilayer. These GPI anchors are cleaved by phosphatidylinositol-specific phospholipases C (PI-PLCs) to release a water-soluble protein with an exposed glycosylinositol moiety and ... More
Non-apoptotic phosphatidylserine externalization induced by engagement of glycosylphosphatidylinositol-anchored proteins.
AuthorsSmrz D, Dráberová L, Dráber P
JournalJ Biol Chem
PubMed ID17284440
The exposure of phosphatidylserine (PS) on the cell surface is a general marker of apoptotic cells. Non-apoptotic PS externalization is induced by several activation stimuli, including engagement of immunoreceptors. Immune cells can also be activated by aggregation of glycosylphosphatidylinositol-anchored proteins (GPI-APs). However, it is unknown whether cell triggering through these ... More
Release of carcinoembryonic antigen from human tumor cells by phosphatidylinositol-specific phospholipase C: highly effective extraction and upregulation from LS-174T colonic adenocarcinoma cells.
AuthorsGouin E, Ouary M, Pogu S, Saï P
JournalArch Biochem Biophys
PubMed ID8215392
Carcinoembryonic antigen (CEA), produced by gastrointestinal tumor cells, is anchored to cell membrane by a glycosyl-phosphatidylinositol moiety which can be cleaved with phosphatidylinositol-specific phospholipase C (PI-PLC). We studied the extraction of CEA from living human colon carcinoma (LS-174T, HT-29, COLO-205, and HRT-18) and pancreatic carcinoma (CAPAN) cells by PI-PLC from ... More
Function-based high-throughput screening for antibody antagonists and agonists against G protein-coupled receptors.
AuthorsRen H, Li J, Zhang N, Hu LA, Ma Y, Tagari P, Xu J, Zhang MY
JournalCommun Biol
PubMed ID32218528
'Hybridoma and phage display are two powerful technologies for isolating target-specific monoclonal antibodies based on the binding. However, for complex membrane proteins, such as G protein-coupled receptors (GPCRs), binding-based screening rarely results in functional antibodies. Here we describe a function-based high-throughput screening method for quickly identifying antibody antagonists and agonists ... More
Hepatic ABCA1 deficiency is associated with delayed apolipoprotein B secretory trafficking and augmented VLDL triglyceride secretion.
AuthorsLiu M, Chung S, Shelness GS, Parks JS
JournalBiochim Biophys Acta Mol Cell Biol Lipids
PubMed ID28694219
'ATP binding cassette transporter A1 (ABCA1) is a membrane transporter that facilitates nascent HDL formation. Tangier disease subjects with complete ABCA1 deficiency have <5% of normal levels of plasma HDL, elevated triglycerides (TGs), and defective vesicular trafficking in fibroblasts and macrophages. Hepatocyte-specific ABCA1 knockout mice (HSKO) have a similar lipid ... More
Quercetin acutely relaxes airway smooth muscle and potentiates β-agonist-induced relaxation via dual phosphodiesterase inhibition of PLCβ and PDE4.
Authors
JournalAm J Physiol Lung Cell Mol Physiol
PubMed ID23873842
Modified Ham test for atypical hemolytic uremic syndrome.
Authors
JournalBlood
PubMed ID25862562
LDLRAD3 is a receptor for Venezuelan equine encephalitis virus.
Authors
JournalNature
PubMed ID33208938
Loss of the N-acetylgalactosamine side chain of the GPI-anchor impairs bone formation and brain functions and accelerates the prion disease pathology.
Authors
JournalJ Biol Chem
PubMed ID35151686
Direct evidence for a new mode of plant defense against insects via a novel polygalacturonase-inhibiting protein expression strategy.
Authors
JournalJ Biol Chem
PubMed ID32611768
Attenuation of airway smooth muscle contractility via flavonol-mediated inhibition of phospholipase-Cβ.
Authors
JournalAm J Physiol Lung Cell Mol Physiol
PubMed ID26773068
Active components of ginger potentiate β-agonist-induced relaxation of airway smooth muscle by modulating cytoskeletal regulatory proteins.
Authors
JournalAm J Respir Cell Mol Biol
PubMed ID23962082
A solution to Nature's haemoglobin knockout: a plasma-accessible carbonic anhydrase catalyses CO2 excretion in Antarctic icefish gills.
Authors
JournalJ Exp Biol
PubMed ID30291156
Identification of a Golgi GPI-N-acetylgalactosamine transferase with tandem transmembrane regions in the catalytic domain.
Authors
JournalNat Commun
PubMed ID29374258
Roles of two types of heparan sulfate clusters in Wnt distribution and signaling in Xenopus.
AuthorsMii Y, Yamamoto T, Takada R, Mizumoto S, Matsuyama M, Yamada S, Takada S, Taira M
JournalNat Commun
PubMed ID29215008
Wnt proteins direct embryonic patterning, but the regulatory basis of their distribution and signal reception remain unclear. Here, we show that endogenous Wnt8 protein is distributed in a graded manner in Xenopus embryo and accumulated on the cell surface in a punctate manner in association with "N-sulfo-rich heparan sulfate (HS)," ... More
IgSF21 promotes differentiation of inhibitory synapses via binding to neurexin2a.
AuthorsTanabe Y, Naito Y, Vasuta C, Lee AK, Soumounou Y, Linhoff MW, Takahashi H
JournalNat Commun
PubMed ID28864826
Coordinated development of excitatory and inhibitory synapses is essential for higher brain function, and impairment in this development is associated with neuropsychiatric disorders. In contrast to the large body of accumulated evidence regarding excitatory synapse development, little is known about synaptic adhesion and organization mechanisms underlying inhibitory synapse development. Through ... More