2011
Interaction of Recombinant Myocilin with the Matricellular Protein SPARC: Functional Implications
Aroca-Aguilar JD, Sánchez-Sánchez F, Ghosh S, Fernández-Navarro A, Coca-Prados M, Escribano J. Interaction of Recombinant Myocilin with the Matricellular Protein SPARC: Functional Implications. Investigative Ophthalmology & Visual Science 2011, 52: 179-189. PMID: 20926826, PMCID: PMC3053273, DOI: 10.1167/iovs.09-4866.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceBlotting, WesternCalcium-Binding ProteinsCell LineChromatography, High Pressure LiquidCytoskeletal ProteinsExtracellular Matrix ProteinsEye ProteinsGene Expression ProfilingGlycoproteinsHumansKidneyMicroscopy, FluorescenceMolecular Sequence DataOsteonectinPolymerase Chain ReactionProtein BindingRecombinant Fusion ProteinsRecombinant ProteinsTwo-Hybrid System TechniquesConceptsHEK 293T cellsSolid-phase binding assaysInteraction of myocilinRecombinant proteinsRecombinant myocilinMatricellular proteinEC domainTerminal olfactomedin domainTwo-hybrid analysisTwo-hybrid systemProtein-protein interactionsFull-length myocilinC-terminal domainN-terminal domainC-terminal regionCalcium binding domainsBinding assaysOlfactomedin domainC-terminal fragmentBinding domainsLinker regionUnknown functionExtracellular glycoproteinIntracellular interactionsSPARC family
2008
Expression and purification of functional recombinant human pigment epithelium-derived factor (PEDF) secreted by the yeast Pichia pastoris
Sánchez-Sánchez F, Aroca-Aguilar JD, Segura I, Ramírez-Castillejo C, Riese HH, Coca-Prados M, Escribano J. Expression and purification of functional recombinant human pigment epithelium-derived factor (PEDF) secreted by the yeast Pichia pastoris. Journal Of Biotechnology 2008, 134: 193-201. PMID: 18282627, DOI: 10.1016/j.jbiotec.2008.01.005.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBlotting, WesternCell LineChlorocebus aethiopsCOS CellsEye ProteinsHumansMicroscopy, FluorescenceNerve Growth FactorsPichiaRecombinant ProteinsSerpinsConceptsPigment epithelium-derived factorEpithelium-derived factorRecombinant pigment epithelium-derived factorRecombinant human pigment epithelium-derived factorHuman pigment epithelium-derived factorCerebellar granule cell survivalPotential therapeutic agentGranule cell survivalFull-length PEDFStem cell self-renewal propertiesCell-based therapiesTherapeutic roleOcular diseasesTherapeutic agentsSelf-renewal propertiesEndothelial cell migrationLiquid chromatographyCell linesRPEDFCell survivalHigh-performance liquid chromatographyCell migrationYeast Pichia pastorisLow pressure liquid chromatographyPichia pastoris
2005
Myocilin Mutations Causing Glaucoma Inhibit the Intracellular Endoproteolytic Cleavage of Myocilin between Amino Acids Arg226 and Ile227 *
Aroca-Aguilar JD, Sánchez-Sánchez F, Ghosh S, Coca-Prados M, Escribano J. Myocilin Mutations Causing Glaucoma Inhibit the Intracellular Endoproteolytic Cleavage of Myocilin between Amino Acids Arg226 and Ile227 *. Journal Of Biological Chemistry 2005, 280: 21043-21051. PMID: 15795224, DOI: 10.1074/jbc.m501340200.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsArginineBlotting, WesternBrefeldin ACattleCell LineComputational BiologyCOS CellsCulture MediaCytoskeletal ProteinsExtracellular Matrix ProteinsEye ProteinsGlaucomaGlycoproteinsGreen Fluorescent ProteinsHumansIsoleucineLeucineMicroscopy, FluorescenceMolecular Sequence DataMutationPeptidesPhenotypeProtein Structure, TertiarySpectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationTransfectionConceptsMyocilin mutationPathogenesis of glaucomaCause of blindnessSevere glaucoma phenotypeHuman aqueous humorAqueous humorOcular tissuesGlaucoma phenotypeNonocular tissuesWestern immunoblot analysisGlaucomaPathogenic mutationsOlfactomedin-like domainEndoproteolytic processingWild-type myocilinCell linesImmunoblot analysisMyocilinEndoproteolytic cleavageLeucine zipper-like domainMutant myocilinNormal roleHuman organsInhibitionTissue
2001
Chloride dependent intracellular pH effects of external ATP in cultured human non-pigmented ciliary body epithelium
Cullinane A, Coca-Prados M, Harvey B. Chloride dependent intracellular pH effects of external ATP in cultured human non-pigmented ciliary body epithelium. Current Eye Research 2001, 23: 443-447. PMID: 12045894, DOI: 10.1076/ceyr.23.6.443.6967.Peer-Reviewed Original ResearchMeSH KeywordsAdenosine TriphosphateAnti-Bacterial AgentsCalciumCells, CulturedChloride-Bicarbonate AntiportersChloridesCiliary BodyFluoresceinsFluorescent DyesHumansHydrogen-Ion ConcentrationMacrolidesMicroscopy, FluorescencePigment Epithelium of EyeProton-Translocating ATPasesReceptors, PurinergicSpectrometry, FluorescenceSuraminUp-RegulationConceptsNon-pigmented ciliary body epitheliumHNPE cellsCiliary body epitheliumAcetoxymethyl ester formBody epitheliumIntracellular calcium levelsCell-permeable calcium chelatorFluorescent probe BCECFReceptor inhibitorsIntracellular calciumPurinergic receptorsCalcium levelsDiisothiocyanatostilbene-2Extracellular adenosineSustained increaseMM suraminInhibitor amilorideExtracellular ATPCalcium chelatorEster formMicroM ATPATPase inhibitorEpitheliumInhibitorsIntracellularExtracellular ATP effects on calcium signaling in cultured human non-pigmented ciliary body epithelium
Cullinane A, Coca-Prados M, Harvey B. Extracellular ATP effects on calcium signaling in cultured human non-pigmented ciliary body epithelium. Current Eye Research 2001, 23: 448-454. PMID: 12045895, DOI: 10.1076/ceyr.23.6.448.6964.Peer-Reviewed Original ResearchMeSH KeywordsAdenosine TriphosphateCalciumCalcium-Transporting ATPasesCells, CulturedCiliary BodyFluorescent DyesFura-2HumansMicroscopy, FluorescencePigment Epithelium of EyeReceptors, Purinergic P2Receptors, Purinergic P2Y2Signal TransductionSodium-Calcium ExchangerSpectrometry, FluorescenceThapsigarginConceptsNon-pigmented ciliary body epitheliumCiliary body epitheliumHNPE cellsIntracellular storesBody epitheliumFluorescent dye fura-2Extracellular ATP effectP2Y2 receptor activationDye fura-2Calcium signalsATPase pumpPresence of thapsigarginCapacitative calcium entrySodium-free solutionIntracellular calciumPurinergic receptorsFura-2Therapeutic targetCalcium entryPotency orderReceptor activationSustained increaseL-typeExtracellular ATPOscillatory increases
2000
Na+/H+ and Cl–/HCO3–-antiporters of bovine pigmented ciliary epithelial cells
Counillon L, Touret N, Bidet M, Peterson-Yantorno K, Coca-Prados M, Stuart-Tilley A, Wilhelm S, Alper S, Civan M. Na+/H+ and Cl–/HCO3–-antiporters of bovine pigmented ciliary epithelial cells. Pflügers Archiv - European Journal Of Physiology 2000, 440: 667-678. PMID: 11007305, DOI: 10.1007/s004240000302.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsAnion Transport ProteinsAntiportersCattleCell Line, TransformedChloride-Bicarbonate AntiportersCiliary BodyEpithelial CellsImmunohistochemistryMembrane ProteinsMicroscopy, FluorescenceMicroscopy, VideoModels, BiologicalPigmentationReverse Transcriptase Polymerase Chain ReactionRNA, MessengerSLC4A ProteinsSodiumSodium-Hydrogen ExchangersConceptsAqueous humor formationCl-/HCO3Epithelial cell layerTranscriptase-polymerase chain reaction amplificationPE cellsCell layerHuman ciliary bodyAE2 Cl-/HCO3Medical therapyNHE-1 isoformCiliary epithelial cellsCiliary bodyAqueous humorCiliary epithelial bilayerPolymerase chain reaction amplificationChain reaction amplificationNHE-1Rabbit ciliary epitheliumCiliary epitheliumEpithelial cellsBovine pigmented ciliary epithelial cellsGap junctionsRegulatory volume increasePigmented ciliary epithelial cellsExternal Cl
1991
Expression of multiple Na+,K+‐ATPase genes reveals a gradient of isoforms along the nonpigmented ciliary epithelium: Functional implications in aqueous humor secretion
Ghosh S, Hernando N, Martín‐Alonso J, Martin‐Vasallo P, Coca‐Prados M. Expression of multiple Na+,K+‐ATPase genes reveals a gradient of isoforms along the nonpigmented ciliary epithelium: Functional implications in aqueous humor secretion. Journal Of Cellular Physiology 1991, 149: 184-194. PMID: 1660898, DOI: 10.1002/jcp.1041490203.Peer-Reviewed Original Research