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The Caveolin3 complex stabilizes functional surface expression of the Lactate-Proton Shuttle McT1 but not the sodium channel Nav1.5 in cardiomyocytes

ABSTRACT: Caveolin3 variants were associated with action potential prolongation. As Caveolin1 was recently identified in cardiomyocytes, we hypothesize that conserved isoform-specific protein interactions underlie human loss-of-function variants. To analyze the Caveolin1 and Caveolin3 interactome, we developed unbiased live-cell proteomic and isoform-specific mass spectrometry techniques. We demonstrate the functional relevance and pathogenic mechanism of a novel Caveolin3 interactor in gene-edited human iPSC-cardiomyocyte models.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Cardiac Muscle Cell

DISEASE(S): Cardiovascular System Disease

SUBMITTER: Christof Lenz

LAB HEAD: Christof Lenz

PROVIDER: PXD019380 | Pride | 2022-10-30

REPOSITORIES: Pride

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Project description:The trophic condition related proteomes of an oxygenic photosynthetic cyanobacterium Synechocystis sp. PCC 6803 (glucose tolerant strain) were analyzed to get further insights into the metabolic traits governing the early exponential growth under photoautotrophic, mixotrophic and heterotrophic conditions. The cells were grown under constant light autotrophically (AT) in low (air-level) CO2 (ATLC) and in high CO2 (ATHC), mixotrophically (MT) in low (air-level) CO2 in the presence of glucose (MTLC) and heterotrophically in low (air-level) CO2 in the presence of glucose and only 10 min exposure to light every 24 hours (Light-Activated Heterotrophic Growth; LAHGLC). The proteomes of ATHC, MTLC and LAHGLC differed substantially from that of ATLC, particularly by downregulation of the inducible carbon concentrating mechanisms (CCM) and photorespiration related enzymes. On the contrary, under carbon-rich conditions (ATHC, MTLC, LAHGLC) the cells accumulated respiratory NAD(P)H dehydrogenase I complexes in the thylakoid membrane. In glucose supplemented cultures (MTLC, LAHGLC) a distinct NDH-2 protein, NdbA, additionally accumulated in the thylakoid membrane whilst the plasma membrane localized NdbC and ARTO decreased in abundance. The light-harvesting proteins together with the photosystem I and II subunits were uniquely depleted under LAHGLC condition, accumulated under ATHC and in MTLC remained at the level of the reference ATLC condition. The accumulation of P-transporters was unique, revealing only minor differences between ATLC and ATHC but strong up-regulation in MTLC and down-regulation in LAHGLC. This likely implies that MTLC stores energy surplus in the highly energetic bonds of polyphosphate polymer that can be used under unfavorable growth conditions. It is concluded that the rigor of cell growth in MTLC condition results to a great extent from high intracellular inorganic carbon conditions created upon glucose breakdown and release of CO2, beside the direct utilization of glucose-derived carbon skeletons for growth. This combination provides the MTLC cultures with excellent conditions for growth that often exceeds that of the mere photoautotrophic growth at high CO2.

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The Caveolin3 complex stabilizes functional surface expression of the Lactate-Proton Shuttle McT1 but not the sodium channel Nav1.5 in cardiomyocytes

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