4-Hydroxynonenal dependent alteration of TRPV1-mediated coronary microvascular signaling

Autor: Pritam Sinharoy, Daniel J. DelloStritto, Holly C. Cappelli, Ian N. Bratz, Derek S. Damron, Werner J. Geldenhuys, Patrick J. Connell, Charles K. Thodeti, Joseph N. Fahmy
Rok vydání: 2016
Předmět:
Male
0301 basic medicine
Patch-Clamp Techniques
Mutant
Action Potentials
Biochemistry
Lipid peroxidation
Mice
chemistry.chemical_compound
chemistry.chemical_classification
musculoskeletal
neural
and ocular physiology
Coronary Vessels
Amino acid
Cell biology
Femoral Artery
Vasodilation
lipids (amino acids
peptides
and proteins)
Blood Flow Velocity
psychological phenomena and processes
Signal Transduction
medicine.medical_specialty
TRPV Cation Channels
Oxidative phosphorylation
Biology
Article
4-Hydroxynonenal
03 medical and health sciences
In vivo
Coronary Circulation
Physiology (medical)
Internal medicine
Diabetes Mellitus
medicine
Animals
Humans
Calcium Signaling
Cysteine
Aldehydes
HEK 293 cells
Cardiovascular Agents
Mice
Inbred C57BL
Disease Models
Animal
HEK293 Cells
030104 developmental biology
Endocrinology
nervous system
chemistry
Lipid Peroxidation
Capsaicin
Protein Processing
Post-Translational
Zdroj: Free Radical Biology and Medicine. 101:10-19
ISSN: 0891-5849
DOI: 10.1016/j.freeradbiomed.2016.09.021
Popis: We demonstrated previously that TRPV1-dependent regulation of coronary blood flow (CBF) is disrupted in diabetes. Further, we have shown that endothelial TRPV1 is differentially regulated, ultimately leading to the inactivation of TRPV1, when exposed to a prolonged pathophysiological oxidative environment. This environment has been shown to increase lipid peroxidation byproducts including 4-Hydroxynonenal (4-HNE). 4-HNE is notorious for producing protein post-translation modification (PTM) via reactions with the amino acids: cysteine, histidine and lysine. Thus, we sought to determine if 4-HNE mediated post-translational modification of TRPV1 could account for dysfunctional TRPV1-mediated signaling observed in diabetes. Our initial studies demonstrate 4-HNE infusion decreases TRPV1-dependent coronary blood flow in C57BKS/J (WT) mice. Further, we found that TRPV1-dependent vasorelaxation was suppressed after 4-HNE treatment in isolated mouse coronary arterioles. Moreover, we demonstrate 4-HNE significantly inhibited TRPV1 currents and Ca2+ entry utilizing patch-clamp electrophysiology and calcium imaging respectively. Using molecular modeling, we identified potential pore cysteines residues that, when mutated, could restore TRPV1 function in the presence of 4-HNE. Specifically, complete rescue of capsaicin-mediated activation of TRPV1 was obtained following mutation of pore Cysteine 621. Finally, His tag pull-down of TRPV1 in HEK cells treated with 4-HNE demonstrated a significant increase in 4-HNE binding to TRPV1, which was reduced in the TRPV1 C621G mutant. Taken together these data suggest that 4-HNE decreases TRPV1-mediated responses, at both the in vivo and in vitro levels and this dysfunction can be rescued via mutation of the pore Cysteine 621. Our results show the first evidence of an amino acid specific modification of TRPV1 by 4-HNE suggesting this 4-HNE-dependent modification of TRPV1 may contribute to microvascular dysfunction and tissue perfusion deficits characteristic of diabetes.
Databáze: OpenAIRE
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