2018
Identification of myocilin as a blood plasma protein and analysis of its role in leukocyte adhesion to endothelial cell monolayers
Aroca-Aguilar JD, Fernández-Navarro A, Ontañón J, Coca-Prados M, Escribano J. Identification of myocilin as a blood plasma protein and analysis of its role in leukocyte adhesion to endothelial cell monolayers. PLOS ONE 2018, 13: e0209364. PMID: 30557320, PMCID: PMC6296516, DOI: 10.1371/journal.pone.0209364.Peer-Reviewed Original ResearchMeSH KeywordsAdultAgedAged, 80 and overBlood ProteinsBlotting, WesternCell AdhesionCytoskeletal ProteinsEye ProteinsFemaleGlycoproteinsHealthy VolunteersHEK293 CellsHuman Umbilical Vein Endothelial CellsHumansLeukocytesLiverMaleMiddle AgedProteolysisReal-Time Polymerase Chain ReactionRNA, MessengerThymus GlandConceptsPresence of myocilinEndothelial cell monolayersWestern immunoblotNon-ocular tissuesCell monolayersLymphoid organsLymphoid tissueT lymphocytesLeukocyte adhesionMatricellular proteinPlasma proteinsHuman myocilinLeukocytesMyocilinSerum proteinsPutative roleQuantitative PCRBlood plasmaLiverBiological activityAnti-adhesive proteinImmunoblotVivo proteolytic processingNew biological propertiesTissue
2013
Bicarbonate-Dependent Secretion and Proteolytic Processing of Recombinant Myocilin
Aroca-Aguilar JD, Martínez-Redondo F, Martín-Gil A, Pintor J, Coca-Prados M, Escribano J. Bicarbonate-Dependent Secretion and Proteolytic Processing of Recombinant Myocilin. PLOS ONE 2013, 8: e54385. PMID: 23342144, PMCID: PMC3547000, DOI: 10.1371/journal.pone.0054385.Peer-Reviewed Original ResearchMeSH KeywordsAqueous HumorBicarbonatesBlotting, WesternCell LineCulture MediaCytoskeletal ProteinsEye ProteinsGlycoproteinsHumansProteolysisConceptsIrreversible visual lossElevated intraocular pressureIntracellular accumulationC-terminal fragmentVisual lossOptic neuropathyIntraocular pressureIntracellular proteolytic processingProteolytic processingOcular tissuesRecombinant myocilinCalpain IICellular modelMyocilinSecretionPossible factorsExtracellular glycoproteinNeuropathyGlaucoma
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
2009
Functional Role of Proteolytic Processing of Recombinant Myocilin in Self-Aggregation
Aroca-Aguilar JD, Martínez-Redondo F, Sánchez-Sánchez F, Coca-Prados M, Escribano J. Functional Role of Proteolytic Processing of Recombinant Myocilin in Self-Aggregation. Investigative Ophthalmology & Visual Science 2009, 51: 72-78. PMID: 19696176, PMCID: PMC2869055, DOI: 10.1167/iovs.09-4118.Peer-Reviewed Original ResearchLocalization of Multidrug Resistance-Associated Protein 2 in the Nonpigmented Ciliary Epithelium of the Eye
Pelis RM, Shahidullah M, Ghosh S, Coca-Prados M, Wright SH, Delamere NA. Localization of Multidrug Resistance-Associated Protein 2 in the Nonpigmented Ciliary Epithelium of the Eye. Journal Of Pharmacology And Experimental Therapeutics 2009, 329: 479-485. PMID: 19201990, PMCID: PMC2672870, DOI: 10.1124/jpet.108.149625.Peer-Reviewed Original ResearchConceptsNonpigmented epitheliumMicroM MK571Ciliary bodyMultidrug resistance associated protein 2Therapeutic drugsMrp2 proteinIntracellular accumulationBlood-aqueous barrierNonpigmented ciliary epitheliumProtein 2Human ciliary bodyMicroM indomethacinMRP inhibitorsIntraocular tissuesAqueous humorApical membraneMRP2 mRNAWestern blotMultidrug resistanceCiliary epitheliumMicroM cyclosporinPorcine eyesMRP2Cell layerNative human
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 ResearchConceptsPigment 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
2007
Molecular Analysis of Neurolysin Expression in the Rat and Bovine Ciliary Body
Bertazolli-Filho R, Coca-Prados M, Haddad A, Laicine EM. Molecular Analysis of Neurolysin Expression in the Rat and Bovine Ciliary Body. Current Eye Research 2007, 32: 751-756. PMID: 17882707, DOI: 10.1080/02713680701573381.Peer-Reviewed Original Research
2005
Retinoid processing proteins in the ocular ciliary epithelium.
Salvador-Silva M, Ghosh S, Bertazolli-Filho R, Boatright JH, Nickerson JM, Garwin GG, Saari JC, Coca-Prados M. Retinoid processing proteins in the ocular ciliary epithelium. Molecular Vision 2005, 11: 356-65. PMID: 15928609.Peer-Reviewed Original ResearchAcyltransferasesAlcohol OxidoreductasesAnimalsATP-Binding Cassette TransportersBlotting, WesternCarrier ProteinsCattleCells, CulturedChromatography, High Pressure LiquidCiliary BodyCis-trans-IsomerasesEye ProteinsFluorescent Antibody Technique, IndirectGene Expression RegulationHumansImmunohistochemistryPigment Epithelium of EyeProtein TransportRabbitsRetinoidsRetinol-Binding ProteinsRetinol-Binding Proteins, CellularReverse Transcriptase Polymerase Chain ReactionRNA, MessengerMyocilin 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
2004
The bovine iris–ciliary epithelium expresses components of rod phototransduction
Ghosh S, Salvador-Silva M, Coca-Prados M. The bovine iris–ciliary epithelium expresses components of rod phototransduction. Neuroscience Letters 2004, 370: 7-12. PMID: 15489008, DOI: 10.1016/j.neulet.2004.07.026.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsAnimals, NewbornArrestinBlotting, NorthernBlotting, WesternCattleCiliary BodyEpitheliumEye ProteinsGene ExpressionG-Protein-Coupled Receptor Kinase 1IrisLight Signal TransductionNeuronsPromoter Regions, GeneticProtein KinasesRetinal Rod Photoreceptor CellsReverse Transcriptase Polymerase Chain ReactionRhodopsinRNA, MessengerTransfectionConceptsCiliary epitheliumIris cellsBasal activityCommon embryonic originOcular ciliary epitheliumNeural retinaWestern blotRT-PCR amplificationRetinaEpitheliumSignificant stimulationBovine irisEmbryonic originStimulationBlotRod phototransductionTransient transfectionNorthern blotRhodopsin kinasePromoter activityDistal promoter elementLow levelsIrisReporter constructsLower vertebrates
2001
Molecular Evidence That Human Ocular Ciliary Epithelium Expresses Components Involved in Phototransduction
Bertazolli-Filho R, Ghosh S, Huang W, Wollmann G, Coca-Prados M. Molecular Evidence That Human Ocular Ciliary Epithelium Expresses Components Involved in Phototransduction. Biochemical And Biophysical Research Communications 2001, 284: 317-325. PMID: 11394879, DOI: 10.1006/bbrc.2001.4970.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsAntibodies, MonoclonalArrestinBlotting, WesternCalcium-Binding ProteinsCattleCell LineCiliary BodyCyclic Nucleotide-Gated Cation ChannelsEpithelial CellsEye ProteinsFluorescent Antibody Technique, IndirectG-Protein-Coupled Receptor Kinase 1HippocalcinHumansImmunoblottingImmunohistochemistryIon ChannelsLight Signal TransductionLipoproteinsNerve Tissue ProteinsPhosphoric Diester HydrolasesProtein KinasesProtein SubunitsRecoverinRetinaReverse Transcriptase Polymerase Chain ReactionRhodopsinRNA, MessengerSequence Analysis, DNATransducinConceptsHuman ocular ciliary epitheliumCiliary epitheliumOcular ciliary epitheliumNPE cellsCiliary epithelial cell linePars plicata regionIntact ciliary epitheliumWestern blot analysisCultured NPE cellsEpithelial cell lineExpression of rhodopsinIndirect immunofluorescenceVisual arrestinHuman retinaMonoclonal antibodiesHuman ciliary epitheliumThree- to fourfoldEpitheliumRT-PCRPhotoreceptor cellsRhodopsin mRNABlot analysisCell linesRetinaRhodopsin kinase
2000
Expression of the TIGR gene in the iris, ciliary body, and trabecular meshwork of the human eye.
Huang W, Jaroszewski J, Ortego J, Escribano J, Coca-Prados M. Expression of the TIGR gene in the iris, ciliary body, and trabecular meshwork of the human eye. Ophthalmic Genetics 2000, 21: 155-69. PMID: 11035548, DOI: 10.1076/1381-6810(200009)21:3;1-z;ft155.Peer-Reviewed Original ResearchConceptsTIGR proteinTranscription/translation systemCanine pancreatic microsomal membranesPancreatic microsomal membranesSitu hybridization experimentsTissue-specific mannerTIGR genePattern of expressionCarboxy-terminus regionMajor protein bandsHybridization experimentsTerminus regionFusion proteinGlycosylation activityMolecular massProteinPNGase FDeglycosylation treatmentProtein bandsMicrosomal membranesTranslation systemO-glycosidaseJuvenile-onset primary open-angle glaucomaGenes
1997
Gene Expression of Proteases and Protease Inhibitors in the Human Ciliary Epithelium and ODM-2 Cells
ORTEGO J, ESCRIBANO J, COCA-PRADOS M. Gene Expression of Proteases and Protease Inhibitors in the Human Ciliary Epithelium and ODM-2 Cells. Experimental Eye Research 1997, 65: 289-299. PMID: 9268597, DOI: 10.1006/exer.1997.0333.Peer-Reviewed Original ResearchMeSH KeywordsAgedAlpha 1-AntitrypsinAlpha-MacroglobulinsBlotting, NorthernBlotting, WesternCarcinogensCathepsin DCathepsin KCathepsinsCell LineCiliary BodyCysteine EndopeptidasesDNA, ComplementaryEndopeptidasesEpithelial CellsGene ExpressionGlycoproteinsHumansMiddle AgedPolymerase Chain ReactionPrecipitin TestsPregnancy ProteinsProtease InhibitorsSerine Proteinase InhibitorsTetradecanoylphorbol AcetateConceptsODM-2 cellsPP5/TFPICiliary epithelial cellsComplementary DNARegulation of transcriptionProteinase inhibitorsEpithelial cellsNon-protein kinase CCathepsin DProtein kinase C activationKinase C activationPattern of expressionCiliary bodyImmunoprecipitation experimentsGene expressionNorthern analysisCultured ciliary epithelial cellsKinase CWestern blot analysisHuman ciliary epitheliumAlpha-phorbol didecanoatePhorbol esterC activationBlot analysisHuman proteinases
1994
Isolation of cDNA clones encoding the 80-kd subunit protein of the human autoantigen Ku (p70/p80) by antisera raised against ciliary processes of human eye donors.
Hong T, Escribano J, Coca-Prados M. Isolation of cDNA clones encoding the 80-kd subunit protein of the human autoantigen Ku (p70/p80) by antisera raised against ciliary processes of human eye donors. Investigative Ophthalmology & Visual Science 1994, 35: 4023-30. PMID: 7960584.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsAntigens, NuclearAutoantigensBlotting, WesternCell LineCiliary BodyClone CellsDNA HelicasesDNA, ComplementaryDNA-Binding ProteinsFluorescent Antibody TechniqueHumansImmunoglobulin GKu AutoantigenMaleMiceMice, Inbred BALB CMiddle AgedMolecular WeightNuclear ProteinsPigment Epithelium of EyeRNA, MessengerTissue DonorsConceptsHuman eye donorsPolymyositis overlap syndromeScleroderma-polymyositis overlap syndromeODM-2 cellsHuman ciliary processesCiliary processesOverlap syndromeEye donorsCiliary epitheliumSpecies-specific immune responseAnti-Ku antibodiesVariety of antigensCell linesEpithelium cell linePlaque-purified clonesCiliary epithelial cell lineOcular ciliary epitheliumWestern blot analysisSubunit proteinsEpithelial cell lineCiliary epithelial cellsImmune responseOcular tissues
1993
Differential Expression of the Cellular Retinaldehyde-binding Protein in Bovine Ciliary Epithelium
Martı̀n-Alonso J, Ghosh S, Hernando N, Crabb J, Coca-Prados M. Differential Expression of the Cellular Retinaldehyde-binding Protein in Bovine Ciliary Epithelium. Experimental Eye Research 1993, 56: 659-669. PMID: 8595808, DOI: 10.1006/exer.1993.1083.Peer-Reviewed Original Research
1992
Isolation of a cDNA encoding a glutathione S-transferase (GST) class-π from the bovine ocular ciliary epithelium
Hernando N, Martin-Alonso J, Ghosh S, Coca-Prados M. Isolation of a cDNA encoding a glutathione S-transferase (GST) class-π from the bovine ocular ciliary epithelium. Experimental Eye Research 1992, 55: 711-718. PMID: 1478280, DOI: 10.1016/0014-4835(92)90175-r.Peer-Reviewed Original Research
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
1990
Cellular distribution and differential gene expression of the three alpha subunit isoforms of the Na,K-ATPase in the ocular ciliary epithelium.
Ghosh S, Freitag A, Martin-Vasallo P, Coca-Prados M. Cellular distribution and differential gene expression of the three alpha subunit isoforms of the Na,K-ATPase in the ocular ciliary epithelium. Journal Of Biological Chemistry 1990, 265: 2935-2940. PMID: 1689295, DOI: 10.1016/s0021-9258(19)39891-6.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBlotting, NorthernBlotting, WesternCattleCell MembraneCell Transformation, ViralCiliary BodyFlow CytometryFluorescent Antibody TechniqueGene ExpressionIsoenzymesMacromolecular SubstancesMicroscopy, ElectronPigment Epithelium of EyeRatsRNASimian virus 40Sodium-Potassium-Exchanging ATPaseConceptsAlpha-subunit isoformsAlpha isoformSubunit isoformsK-ATPaseBasolateral plasma membrane domainsK-ATPase alpha subunitPlasma membrane domainsImmunoreactive signalPE cellsDifferential gene expressionAlpha 3Alpha 3 polypeptidesNPE cellsCell linesAlpha 2Alpha 1 polypeptideAlpha 1Pattern of expressionAlpha 3 isoformAlpha 1 isoformMembrane domainsSpecific cDNA probesSimian virus 40Differential immunostaining patternAlpha 2 isoform
1989
Expression of Na,K‐ATPase alpha subunit isoforms in the human ciliary body and cultured ciliary epithelial cells
Martin‐Vasallo P, Ghosh S, Coca‐Prados M. Expression of Na,K‐ATPase alpha subunit isoforms in the human ciliary body and cultured ciliary epithelial cells. Journal Of Cellular Physiology 1989, 141: 243-252. PMID: 2553750, DOI: 10.1002/jcp.1041410203.Peer-Reviewed Original ResearchConceptsK-ATPase alpha subunit isoformsAlpha-subunit isoformsSubunit isoformsCultured cellsK-ATPase alpha subunitPE cellsExpression of NaBasolateral membrane domainsNorthern hybridization analysisNPE cell linesAlpha 2 mRNACell linesAlpha 3 isoformMembrane domainsAlpha 3 mRNAAlpha 1Alpha subunitCultured ciliary epithelial cellsWestern blot analysisHybridization analysisIsoformsCiliary epithelial cellsNPE layerBlot analysisK-ATPase