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 ResearchConceptsIrreversible visual lossElevated intraocular pressureIntracellular accumulationC-terminal fragmentVisual lossOptic neuropathyIntraocular pressureIntracellular proteolytic processingProteolytic processingOcular tissuesRecombinant myocilinCalpain IICellular modelMyocilinSecretionPossible factorsExtracellular glycoproteinNeuropathyGlaucoma
2012
The Stoichiometric Transition from Zn6Cu1-Metallothionein to Zn7-Metallothionein Underlies the Up-regulation of Metallothionein (MT) Expression QUANTITATIVE ANALYSIS OF MT-METAL LOAD IN EYE CELLS*
Alvarez L, Gonzalez-Iglesias H, Garcia M, Ghosh S, Sanz-Medel A, Coca-Prados M. The Stoichiometric Transition from Zn6Cu1-Metallothionein to Zn7-Metallothionein Underlies the Up-regulation of Metallothionein (MT) Expression QUANTITATIVE ANALYSIS OF MT-METAL LOAD IN EYE CELLS*. Journal Of Biological Chemistry 2012, 287: 28456-28469. PMID: 22722935, PMCID: PMC3436589, DOI: 10.1074/jbc.m112.365015.Peer-Reviewed Original ResearchConceptsInflammatory eye diseaseRetinal ganglion cellsPresence of TNFαCorneal epithelial cell lineInflammatory cytokinesProinflammatory cytokinesEpithelial cell lineSynthesis of metallothioneinGanglion cellsEye diseaseLevels of metallothioneinCorneal tissueCombination of zincEndothelial cellsCytokinesMT1 isoformsIL1αEffect of zincGene expressionCadaver eyesConcentration of metallothioneinEye cellsMicroarray-based analysisZinc exposureMT isoforms
2011
WDR36 and P53 Gene Variants and Susceptibility to Primary Open-Angle Glaucoma: Analysis of Gene-Gene Interactions
Blanco-Marchite C, Sánchez-Sánchez F, López-Garrido MP, Iñigez-de-Onzoño M, López-Martínez F, López-Sánchez E, Alvarez L, Rodríguez-Calvo PP, Méndez-Hernández C, Fernández-Vega L, García-Sánchez J, Coca-Prados M, García-Feijoo J, Escribano J. WDR36 and P53 Gene Variants and Susceptibility to Primary Open-Angle Glaucoma: Analysis of Gene-Gene Interactions. Investigative Ophthalmology & Visual Science 2011, 52: 8467-8478. PMID: 21931130, PMCID: PMC3208188, DOI: 10.1167/iovs.11-7489.Peer-Reviewed Original ResearchAgedAmino Acid SequenceBase SequenceCase-Control StudiesChromatography, High Pressure LiquidDNA Mutational AnalysisEpistasis, GeneticEye ProteinsFemaleGenetic Predisposition to DiseaseGenetic VariationGenotypeGlaucoma, Open-AngleHumansIntraocular PressureMaleMiddle AgedMolecular Sequence DataOcular HypertensionPolymerase Chain ReactionPolymorphism, Single NucleotideRegulatory Sequences, Nucleic AcidRisk FactorsSequence Analysis, DNATumor Suppressor Protein p53Interaction 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 Research
2008
Heterozygous expression of myocilin glaucoma mutants increases secretion of the mutant forms and reduces extracellular processed myocilin.
Aroca-Aguilar JD, Sánchez-Sánchez F, Martínez-Redondo F, Coca-Prados M, Escribano J. Heterozygous expression of myocilin glaucoma mutants increases secretion of the mutant forms and reduces extracellular processed myocilin. Molecular Vision 2008, 14: 2097-108. PMID: 19023451, PMCID: PMC2585175.Peer-Reviewed Original ResearchConceptsWild-type myocilinWild-type proteinMyocilin mutantsMutant myocilinMutant formsProteolytic processingMissense mutant formsHEK 293T cellsMyocilin geneMutant proteinsSecretory pathwayUnidentified functionExtracellular proteinsMutantsEndoproteolytic processingRecombinant mutantsMyocilin secretionCellular fractionsHeterozygous expressionMyocilinProteinUnknown mechanismExtracellular amountSDS-PAGEHeterozygous stateExpression 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
Characterization of the Intracellular Proteolytic Cleavage of Myocilin and Identification of Calpain II as a Myocilin-processing Protease*
Sánchez-Sánchez F, Martínez-Redondo F, Aroca-Aguilar JD, Coca-Prados M, Escribano J. Characterization of the Intracellular Proteolytic Cleavage of Myocilin and Identification of Calpain II as a Myocilin-processing Protease*. Journal Of Biological Chemistry 2007, 282: 27810-27824. PMID: 17650508, DOI: 10.1074/jbc.m609608200.Peer-Reviewed Original ResearchConceptsExtracellular calciumCalpain IICalcium-activated proteaseIntraocular pressureT cellsIntracellular proteolytic cleavageCalpain inhibitorsCalcium uptakeProteolytic cleavageCalpain inhibitor IVOlfactomedin-like domainCalpain IInhibitor IVMyocilinEndoplasmic reticulumIntracellular processingLumenRNA interference knockdownCalciumProteolytic processingCellsCulture mediumGlaucomaSubcellular fractionationEndoproteolytic processingRole of MYOC and OPTN sequence variations in Spanish patients with primary open-angle glaucoma.
López-Martínez F, López-Garrido M, Sánchez-Sánchez F, Campos-Mollo E, Coca-Prados M, Escribano J. Role of MYOC and OPTN sequence variations in Spanish patients with primary open-angle glaucoma. Molecular Vision 2007, 13: 862-72. PMID: 17615537, PMCID: PMC2770203.Peer-Reviewed Original ResearchMeSH KeywordsAge of OnsetAmino Acid SequenceCell Cycle ProteinsCell LineCytoskeletal ProteinsExonsEye ProteinsFemaleGene FrequencyGenome, HumanGlaucoma, Open-AngleGlycoproteinsHaplotypesHumansLinkage DisequilibriumMaleMembrane Transport ProteinsMiddle AgedMolecular Sequence DataMutationOcular HypertensionOpen Reading FramesPhenotypePolymorphism, Single-Stranded ConformationalPromoter Regions, GeneticSpainTranscription Factor TFIIIAWhite PeopleConceptsPrimary open-angle glaucomaOcular hypertensionOpen-angle glaucomaPOAG patientsSpanish patientsSequence variationExons of myocilinHeterozygous disease-causing mutationsPathogenic mutationsSingle nucleotide polymorphismsPromoter regionRole of myocilinDNA sequence variationPolymorphic GT microsatelliteCommon single nucleotide polymorphismsPCR-DNA sequencingT cellsComplete coding regionPatientsUnrelated patientsMYOC geneOPTN geneGlaucomaDisease-causing mutationsGT microsatelliteNew perspectives in aqueous humor secretion and in glaucoma: The ciliary body as a multifunctional neuroendocrine gland
Coca-Prados M, Escribano J. New perspectives in aqueous humor secretion and in glaucoma: The ciliary body as a multifunctional neuroendocrine gland. Progress In Retinal And Eye Research 2007, 26: 239-262. PMID: 17321191, DOI: 10.1016/j.preteyeres.2007.01.002.Peer-Reviewed Original ResearchConceptsIntraocular pressureCiliary bodyAqueous humor secretionAqueous humorCiliary epitheliumGlaucoma-associated genesImmune privilege statusDevelopment of glaucomaGlutamate-metabolizing enzymesPathophysiology of glaucomaAnti-angiogenic factorsSteroid-converting enzymesDiurnal circadian rhythmCiliary blood flowOcular ciliary bodyAnterior segmentBlood flowMultifactorial eventGlaucomaNeuroendocrine systemOutflow pathwayEndocrine communicationEndocrine systemEndocrine peptidesRegulatory peptides
2005
Interaction of myocilin with the C-terminal region of hevin
Li Y, Aroca-Aguilar JD, Ghosh S, Sánchez-Sánchez F, Escribano J, Coca-Prados M. Interaction of myocilin with the C-terminal region of hevin. Biochemical And Biophysical Research Communications 2005, 339: 797-804. PMID: 16316624, DOI: 10.1016/j.bbrc.2005.11.082.Peer-Reviewed Original ResearchRetinoid 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
2002
Bioinformatics and reanalysis of subtracted expressed sequence tags from the human ciliary body: Identification of novel biological functions.
Escribano J, Coca-Prados M. Bioinformatics and reanalysis of subtracted expressed sequence tags from the human ciliary body: Identification of novel biological functions. Molecular Vision 2002, 8: 315-32. PMID: 12355061.Peer-Reviewed Original ResearchMeSH KeywordsCiliary BodyComputational BiologyExpressed Sequence TagsEye ProteinsGene ExpressionHumansSequence Analysis, DNAConceptsSequence tagsUnidentified sequencesBLAST serviceDNA sequence comparisonsCell cycle regulationNon-coding regionsNon-redundant databaseBLAST search programNovel biological functionNovel candidate genesNovel physiological functionHuman Genome ProjectIndependent ESTsCircadian rhythm controlSubtractive hybridizationCellular signalingCycle regulationSequence comparisonAdditional genesDNA sequencesCytoskeleton structurePDB sequencesGenome ProjectCandidate genesBiological functionsAdult-Onset Primary Open-Angle Glaucoma Caused by Mutations in Optineurin
Rezaie T, Child A, Hitchings R, Brice G, Miller L, Coca-Prados M, Héon E, Krupin T, Ritch R, Kreutzer D, Crick RP, Sarfarazi M. Adult-Onset Primary Open-Angle Glaucoma Caused by Mutations in Optineurin. Science 2002, 295: 1077-1079. PMID: 11834836, DOI: 10.1126/science.1066901.Peer-Reviewed Original ResearchMeSH KeywordsAdultAlternative SplicingAmino Acid SequenceBrainCell Cycle ProteinsChromosome MappingChromosomes, Human, Pair 10Ciliary BodyExonsEye ProteinsFemaleGlaucoma, Open-AngleGolgi ApparatusHeterozygoteHumansIntraocular PressureMaleMembrane Transport ProteinsMiddle AgedMutationMutation, MissenseNerve Tissue ProteinsOcular HypertensionPedigreePolymorphism, Single-Stranded ConformationalRetinaTrabecular MeshworkTranscription Factor TFIIIAZinc FingersConceptsPrimary open-angle glaucomaHereditary primary open-angle glaucomaNormal intraocular pressureOpen-angle glaucomaAdult-onset primary open-angle glaucomaNeuroprotective roleIntraocular pressureAngle glaucomaTumor necrosisLeading causeTrabecular meshworkOPTN geneCiliary epitheliumCausative genesGlaucomaChromosome 10p14OptineurinSequence alterationsNecrosisRetinaIndividualsBrainEpithelium
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
Cloning and characterization of subtracted cDNAs from a human ciliary body library encoding TIGR, a protein involved in juvenile open angle glaucoma with homology to myosin and olfactomedin
Ortego J, Escribano J, Coca-Prados M. Cloning and characterization of subtracted cDNAs from a human ciliary body library encoding TIGR, a protein involved in juvenile open angle glaucoma with homology to myosin and olfactomedin. FEBS Letters 1997, 413: 349-353. PMID: 9280311, DOI: 10.1016/s0014-5793(97)00934-4.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceBase SequenceCiliary BodyCloning, MolecularCytoskeletal ProteinsDNA, ComplementaryExtracellular Matrix ProteinsEyeEye ProteinsGene LibraryGlaucoma, Open-AngleGlycoproteinsHumansMolecular Sequence DataMyosinsOrgan SpecificityRNA, MessengerSequence Homology, Amino AcidSequence Homology, Nucleic Acid