Proteomic Landscape and Deduced Functions of the Cardiac 14-3-3 Protein Interactome

Abstract

Rationale: The 14-3-3 protein family is known to interact with many proteins in non-cardiac cell types to regulate multiple signaling pathways, particularly those relating to energy and protein homeostasis; and the 14-3-3 network is a therapeutic target of critical metabolic and proteostatic signaling in cancer and neurological diseases. Although the heart is critically sensitive to nutrient and energy alterations, and multiple signaling pathways coordinate to maintain the cardiac cell homeostasis, neither the structure of cardiac 14-3-3 protein interactome, nor potential functional roles of 14-3-3 protein–protein interactions (PPIs) in heart has been explored. Objective: To establish the comprehensive landscape and characterize the functional role of cardiac 14-3-3 PPIs. Methods and Results: We evaluated both RNA expression and protein abundance of 14-3-3 isoforms in mouse heart, followed by co-immunoprecipitation of 14-3-3 proteins and mass spectrometry in left ventricle. We identified 52 proteins comprising the cardiac 14-3-3 interactome. Multiple bioinformatic analyses indicated that more than half of the proteins bound to 14-3-3 are related to mitochondria; and the deduced functions of the mitochondrial 14-3-3 network are to regulate cardiac ATP production via interactions with mitochondrial inner membrane proteins, especially those in mitochondrial complex I. Binding to ribosomal proteins, 14-3-3 proteins likely coordinate protein synthesis and protein quality control. Localizations of 14-3-3 proteins to mitochondria and ribosome were validated via immunofluorescence assays. The deduced function of cardiac 14-3-3 PPIs is to regulate cardiac metabolic homeostasis and proteostasis. Conclusions: Thus, the cardiac 14-3-3 interactome may be a potential therapeutic target in cardiovascular metabolic and proteostatic disease states, as it already is in cancer therapy.

Shared Proteins and Pathways of Cardiovascular and Cognitive Diseases: Relation to Vascular Cognitive Impairment

“…Of the proteins found crucial only based on Global Network analysis, SOD2, JAK2, APOE, , ALDOA, VCAM1, AKT1, APP, PSEN1, NEDD4, and PRKN were previously already related to VCI. Relations to both CD and CVD were found for MAPK14, , YWHAZ, , CREB3, , HSPB1, , and PRDX6; , however, a direct relation to VCI was not found previously. SOD2, also known as manganese (Mn)-dependent superoxide dismutase, is located in the mitochondrial matrix and is an enzyme protective against OS .…”

Reprogramming of cardiac phosphoproteome, proteome and transcriptome confers resilience to chronic adenylyl cyclase-driven stress

“…Chronic high level of AC8 activation elicits numerous stress responses (8) including increased nutrient sensing with increase AMPK signaling (10A), increased ROS scavenging, protection against cell death, and also induction of signaling to minimize the impact of AC-PKA signaling (10B). Shifts in energy production and utilization to deal with the marked energy demands of the TGAC8 heart (10,12) include: enhancement of glucose metabolism is pronounced (11,13), whilst utilization of fatty acids to produce energy is reduced (11) but an increase in glucose catabolism (10,11) is accompanied by an increased anabolism (12), supported by increased aerobic glycolysis (11,13); increased utilization of the pentose phosphate shunt (5,13) supports increased amino acid, nucleotide, protein and lipoprotein synthesis induced by growth factor signaling (5), resulting in an increase in LV biomass (14). This adaptive profile balances energy requirements necessary for an incessant marked increase in cardiac performance (4), for increased RNA transcription, protein translation and quality control required for cardiac growth (14), and for other biological processes within the LV myocardium (6).…”

“…This adaptive profile balances energy requirements necessary for an incessant marked increase in cardiac performance (4), for increased RNA transcription, protein translation and quality control required for cardiac growth (14), and for other biological processes within the LV myocardium (6). This pattern of consilient adaptations (6)(7)(8)(9)(10)(11)(12)(13)(14)(15) sustains an incessant increase in cardiac performance while conferring cardio-protection to the TGAC8 heart.…”

“…As the body is a quite complex system, regulation at a single level cannot reveal the whole picture in an organism, and multiple levels of regulatory mechanisms are coordinated to defend against stimuli and maintain health 11 . Phosphorylation is one of the most important and most extensively studied post-translational modifications (PTMs) on proteins.…”

“…As the body is a complex system, multiple levels of coordinated regulatory mechanisms are requisite to defend against stimuli and maintain health [6][7][8][9] . Phosphorylation is one of the most important and has been the most extensively studied post-translational modifications (PTM) of proteins.…”