This project focuses on a new cardiac signaling pathway by which mitochondria regulate cytoplasmic regulatory proteins and surface membrane turnover. The pathway relies on the accumulation by mitochondria of coenzyme A (CoA) to a high concentration, such that free CoA gradients to the cytoplasm are greater than 50 to1. For this reason, transient openings of nonselective permeability transition pores (PTP's) in the inner mitochondrial membrane can generate micromolar CoA transients in the cytoplasm without significantly depleting other mitochondrial substrates. CoA transients are then converted to acyl CoA transients because acyl CoA sythetases are limited by the prevailing free cytoplasmic CoA concentration. Numerous signaling proteins will be affected. In extreme metabolic stress, as occurs upon reoxygenation of ischemic cardiac tissue, palmitoylation of surface membrane proteins via this pathway evidently leads to their clustering in liquid ordered (Lo) membrane domains followed by their internalization as massive endocytosis (pMEND). In this context pMEND is detrimental, but preliminary data indicate that the pMEND pathway contributes to constitutive sarcolemma turnover and regulates the activities of Na/K pumps in cardiac myocytes. Using multiple mice lines with deficiencies in this pathway, as well as drugs to block PTP's, we will determine what physiological and pathological roles pMEND-related endocytosis plays in cardiac myocytes. The project will provide insight into fundamental cell regulatory mechanisms that have a high impact for an understanding of the leading cause of death in the developed world.

Public Health Relevance

We recently discovered a cardiac signaling pathway in which mitochondria release a metabolite, coenzyme A, that is used to activate cytoplasmic fatty acids that are then linked to membrane proteins by a process called palmitoylation. At the cell surface, this pathway leads to the removal of proteins into membrane vesicles, and in the wake of ischemic events in the heart large fractions of the cell surface can be removed. We will now determine how this pathway is regulated, is used by, and affects heart cells both in normal physiology and pathological circumstances close to the threshold of death.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL119843-01A1
Application #
8698126
Study Section
Electrical Signaling, Ion Transport, and Arrhythmias Study Section (ESTA)
Program Officer
Wang, Lan-Hsiang
Project Start
2014-04-01
Project End
2018-03-31
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
1
Fiscal Year
2014
Total Cost
$397,500
Indirect Cost
$147,500
Name
University of Texas Sw Medical Center Dallas
Department
Physiology
Type
Schools of Medicine
DUNS #
800771545
City
Dallas
State
TX
Country
United States
Zip Code
75390
Hilgemann, Donald W; Dai, Gucan; Collins, Anthony et al. (2018) Lipid signaling to membrane proteins: From second messengers to membrane domains and adapter-free endocytosis. J Gen Physiol 150:211-224
Schmiege, Philip; Fine, Michael; Li, Xiaochun (2018) The regulatory mechanism of mammalian TRPMLs revealed by cryo-EM. FEBS J 285:2579-2585
Lu, Fang-Min; Hilgemann, Donald W (2017) Na/K pump inactivation, subsarcolemmal Na measurements, and cytoplasmic ion turnover kinetics contradict restricted Na spaces in murine cardiac myocytes. J Gen Physiol 149:727-749
Dalton, George; An, Sung-Wan; Al-Juboori, Saif I et al. (2017) Soluble klotho binds monosialoganglioside to regulate membrane microdomains and growth factor signaling. Proc Natl Acad Sci U S A 114:752-757
Schmiege, Philip; Fine, Michael; Blobel, Günter et al. (2017) Human TRPML1 channel structures in open and closed conformations. Nature 550:366-370
Lu, Fang-Min; Deisl, Christine; Hilgemann, Donald W (2016) Profound regulation of Na/K pump activity by transient elevations of cytoplasmic calcium in murine cardiac myocytes. Elife 5:
Reilly, Louise; Howie, Jacqueline; Wypijewski, Krzysztof et al. (2015) Palmitoylation of the Na/Ca exchanger cytoplasmic loop controls its inactivation and internalization during stress signaling. FASEB J 29:4532-43
Hilgemann, Donald W (2014) Cardiac electrophysiology delivered a ""grand slam"" by angiotensin II: the third explanation of transmural cardiac electrical activity gradients. Biophys J 106:2288-90