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Polat, O. K.

Publications and source records attributed to Polat, O. K..

2 recordsLinked to original sources

Atomistic Insights into TRPC6/Caveolin-1 Interactions Interface via All- Atom Molecular Dynamics Simulations: Structural and Energetic Basis for Selective Modulation

The molecular determinants governing TRPC6 stabilization and its interaction with caveolin1 (CAV1) remain poorly defined, despite their critical role in caveolae organization and signaling. The absence of atomistic structural models has hindered a mechanistic understanding of how TRPC6 is recruited to, and stabilized within, caveolar microdomains. Here, we combine all-atom molecular dynamics simulations with MM/PBSA calculations to characterize the TRPC6-CAV1 complex at atomic resolution. To preserve a biologically realistic membrane environment, harmonic restraints were applied to transmembrane and membrane-embedded regions of both proteins, while the cytosolic TRPC6 N-terminus and solvent-exposed edges of the CAV1 scaffolding domain were kept fully flexible, allowing the putative caveolin-binding motif to explore conformational space and form dynamic contacts. This protocol maintained overall structural integrity while capturing physiologically relevant flexibility at the interaction surface. MM/PBSA analysis revealed a highly favorable binding free energy ({Delta}G-binding = -255.9 {+/-} 1.6 kJ/mol), dominated by electrostatic contributions and reinforced by hydrophobic and aromatic interactions. Per-residue energy decomposition identified an acidic patch in TRPC6 (residues 30-41) that engages a complementary basic, amphipathic segment in CAV1 (residues 85-106), defining a cooperative, reversible binding interface. These findings provide the first atomistic description of TRPC6 recruitment by CAV1 and establish a quantitative framework for the rational design of strategies to selectively modulate this interaction.

biophysics↗

Evaluation of podocyte Rac-1 induced kidney disease by modulation of TRPC5

BackgroundTransient receptor potential channel 5 (TRPC5) is a non-selective cationic ion channel expressed in brain, kidney and other organs where its activation underlies podocyte injury in chronic kidney diseases. Specifically, it has been suggested that a podocyte TRPC5 plasma membrane relocation and channel activation following injury results from activation of Rac-1, propagating podocyte dysfunction and proteinuria. However, previous TRPC5 transgenic mouse studies had questioned a pathogenic role for TRPC5 in podocytes. This investigation was designed to specifically evaluate podocyte Rac-1 activation in the context of functional TRPC5 or a TRPC5 pore mutant to assess effects on proteinuria. Materials and MethodsWe employed single cell patch-clamp studies of cultured podocytes and studied proteinuria in transgenic mouse models to characterize the effects of TRPC5 following podocyte Rac-1 activation. ResultsInhibition of TRPC5 by small molecules reportedly ameliorated proteinuria in murine models of proteinuric kidney diseases. In order to directly examine TRPC5 function following Rac-1-induced podocyte injury, we analyzed TRPC5 inhibition in podocyte specific Rac-1 (active) transgenic mice. In addition, we generated a double-transgenic mouse constitutively overexpressing either TRPC5 (TRPC5WT) or a TRPC5 dominant-negative pore mutant (TRPC5DN) in concert with podocyte specific and inducible activation of active Rac-1 (Rac-1Dtg). In electrophysiological experiments, active TRPC5 was detected in primary podocytes overexpressing TRPC5 but not in podocytes with endogenous TRPC5 expression, nor with Rac-1 overexpressing podocytes. TRPC5 inhibition did not change proteinuria in mice with active podocyte Rac-1, nor did an increase or loss of TRPC5 activity affected podocyte injury in Rac-1Dtg animals. Administration of TRPC5 inhibitors, ML204 and AC1903, did not alleviate podocyte Rac-1 induced proteinuria. ConclusionTRPC5 inhibition did not modify podocyte Rac-1 induced proteinuria in mice. Significance StatementTRPC5 is a calcium conducting ion channel involved in a plethora of biological functions in the brain, kidney and other organs. In proteinuric kidney diseases, others proposed a model that links activation of small GTPase Rac-1 in podocytes to activation of TRPC5 channels propagating cellular injury and eventually leading to progressive kidney disease. To test this hypothesis, we have developed a novel transgenic mouse model that employs podocyte Rac-1 activation in the presence or absence of a functional TRPC5 channel. Our data shows that transgenic mice with activated Rac-1 in podocytes did not enhance endogenous TRPC5 expression or its activity. Furthermore, TRPC5 blockade or activation did not modify Rac-1 induced proteinuria in mice.

cell biology↗