Molecular Characterization of Autophagic Responses Part B

Molecular Characterization of Autophagic Responses, Part B presents a collection of methods for the qualitative and quantitative evaluation of virtually all the morphological, biochemical, and functional manifestations of autophagy, in vitro, ex vivo and in vivo, in organisms as distant as yeast and man. Autophagy is an evolutionarily conserved mechanism for the lysosomal degradation of superfluous or dangerous cytoplasmic entities, and plays a critical role in the preservation of cellular and organismal homeostasis. Monitoring the biochemical processes that accompany autophagy is fundamental for understanding whether autophagic responses are efficient or dysfunctional. Offers a detailed overview of the protocols used to study autophagy and various aspects of autophagic responsesWritten in an accessible style by renowned experts in the field INDICE: Dynamics of autophagosome formation in living cellsNicholas T. KtistakisQuantifying autophagic structures in mammalian cells using confocal microscopySharon A. ToozeDequenching assays for autophagosomal turnoverKathleen Boesze-BattagliaTurnover of lipidated LC3 and autophagic cargoes in mammalian cells and tissuesJosé Manuel FuentesHigh-throughput quantification of GFP-LC3+ dots by automated fluorescence microscopyJosé Manuel Bravo-San PedroUse of pHlurorin-mKate2-LC3 to monitor auophagic responsesIsei TanidaProduction of human ATG proteins for lipidation assaysBrenda A. SchulmanStructural characterization of proteins from the LC3 familyDieter WillboldMethods for studying interactions between LC3/GABARAPs and LIR-containing proteinsVladimir KirkinAssessment of post-translational modifications of ATG proteinsDaolin TangTagged ATG8-coding constructs for the in vivo assessment of ATG4 activityCarlos Lopez-Otin and Guillermo MariñoMeasurement of the Activity of the ATG4 Cysteine ProteasesXiao-Ming YinCristallographic characterization of ATG proteins and interacting partnersBrenda A. SchulmanDynamics of ATG5-ATG12-ATG16L1 aggregation and deaggregationTheocharis PanaretakisFluorescent FYVE chimeras to quantify PtdIns3p synthesis during autophagyMireia Niso-SantanoQuantification of phosphatidylinositol phosphate species in purified membranesAlbert HaasMass assays to quantify bioactive PtdIns3P and PtdIns5P during autophagic responses Bernard PayrastreCellular assays to analyse PtdIns3P and PtdIns5P in autophagyDavid C. RubinszteinUltrastuctural characterization of autophagosomes and autolysosomesEeva-Liisa EskelinenA simple cargo sequestration assay for measurement of non-selective macroautophagy in cultured cellsNikolai Engedal, Morten Luhr, Paula Szalai, Frank Sætre, Lisa Gerner and Per O. Seglen In vitro reconstitution of autophagosome-lysosome fusionQing ZhongIn vitro reconstitution of LC3 lipidationSascha MartensStudy of ULK1 catalytic activity and its regulationBjörn StorkAssessment of MTORC1 enzymatic functionsKen InokiMethods to study the BECN1 interactome in the course of autophagic responsesGian Maria FimiaIn vitro characterization of VPS34 lipid kinase inhibition by small moleculesBenoit PasquierMethods to detect lysosomal AMPK activationShengcai LinAllosteric modulation of AMPK enzymatic activity: in vitro characterizationJessica Ward and Allan Reyes

• ISBN: 978-0-12-809674-1
• Editorial: Academic Press
• Encuadernacion: Cartoné
• Páginas: 412
• Fecha Publicación: 10/03/2017
• Nº Volúmenes: 1
• Idioma: Inglés