Detecting Cardiovascular Protein-Protein Interactions by Proximity Proteomics

Abstract

Rapidly changing and transient protein-protein interactions regulate dynamic cellular processes in the cardiovascular system. Traditional methods, including affinity purification and mass spectrometry, have revealed many macromolecular complexes in cardiomyocytes and the vasculature. Yet these methods often fail to identify in vivo or transient protein-protein interactions. To capture these interactions in living cells and animals with subsequent mass spectrometry identification, enzyme-catalyzed proximity labeling techniques have been developed in the past decade. Although the application of this methodology to cardiovascular research is still in its infancy, the field is developing rapidly, and the promise is substantial. In this review, we outline important concepts and discuss how proximity proteomics has been applied to study physiological and pathophysiological processes relevant to the cardiovascular system.

Rad regulation of CaV1.2 channels controls cardiac fight-or-flight response

“…Proximity labeling was performed 51 with minor modifications 15,52 . Isolated ventricular cardiomyocytes from mice expressing rabbit α 1C -APEX2 were incubated in labeling solution with 0.5 μM biotinphenol (Iris-biotech) for 30 min.…”

Multi‐omic analyses and network biology in cardiovascular disease

“…However, although AP‐MS approaches are routinely utilized for generating PPI networks, they are limited by the inability to detect in vivo the rapidly changing and transient PPIs responsible for regulating dynamic processes in the cardiovascular system. In contrast, proximity ligation proteomics techniques (e.g., BioID, TurboID) are designed to enable the enzymatic biotinylation of proteins that are in close proximity to a bait protein tagged with a biotin ligase enzyme (i.e., BirA and recombinant derivatives) expressed in living cells, allowing for the detection of both stable and transient PPIs in the native cellular environment [135]. In cardiovascular pathobiology, these techniques have identified interactomes of Kir2.1 pathologically relevant mutants [136], PPIs at the cardiac dyad [137, 138], as well as N ‐cadherin [139], caveolin 3 [140], and phospholamban [141] interacting proteins in cardiomyocytes.…”

Harnessing human genetics and stem cells for precision cardiovascular medicine

“… 35 , 36 , 37 Proximity proteomics captures a wide breadth of transient, dynamic, organelle-specific processes as they exist in the native cellular environment. 38 , 39 , 40 , 41 Its recent application to iPSC-CMs and murine hearts has made it possible to describe physiological and pathological processes vital to the cardiovascular system, 42 including the following: the elucidation of the protein networks spanning between the sarcoplasmic reticulum and transverse tubules, 43 transcription factor interactions in cardiogenesis, 44 , 45 and isoform-specific caveolin interactions in CMs. 46 …”