Section of Molecular and Clinical Pharmacology (Nikolai M. Soldatov, Ph.D.) The long-term objective of our research is to pursue the study of the structure-functional relationships of human cardiac and vascular L-type (Cav1.2) Ca2+ channels, and to investigate molecular correlates for the channel regulation associated with alternative splicing, naturally occurring mutations, and voltage-gated conformational rearrangements. We hypothesize that Ca2+-induced inactivation of the a1C channel is mediated by interactions of identified Ca2+ sensors (Soldatov et al., 1998) with site(s) associated with the pore. The Ca2+ sensors appear to be differently targeted by calmodulin and permeating Ca2+ (Romanin et al., 2000) and contribute to the voltage- and Ca2+-dependent inactivation of the channel. Ca2+ sensors are also crucial for the run-down (Kepplinger et al., 2000a) and modulation by auxiliary beta subunits (Kobrinsky et al., 2004, 2005). We extended our investigation of the molecular mechanisms of inactivation (Abernethy and Soldatov, 2002; Morad and Soldatov, 2005) and identified (a) the molecular determinant of slow voltage-dependent inactivation as a ring of hydrophobic residues in the inner pore region (Shi and Soldatov, 2002), (b) link between slow inactivation and beta-subunit modulation (Kobrinsky et al., 2004), (c) roles of the a1C carboxyl- and amino-termini in inactivation (Kobrinsky et al., 2005). Given the importance of the L-type Ca2+ channel in cardiovascular physiology, we extend our investigation of the human a1C splice variants and pursue the following specific aims: (1) using fluorescence resonance energy transfer (FRET) as molecular ruler, to investigate the voltage-gated molecular rearrangements supporting inactivation of the Ca2+ channel and Ca2+ signal transduction mechanisms. (2) Cloning of the new human cardiac small beta2 subunits (Harry et al., 2004) opens new avenue for the identificatiion of the minimum essential determinant of Ca2+ channels modulation and its small biologically active homologues. (3) Using laser capture microdissection technique, RT-PCR, microarray and MS analysis, we will further investigate the diversity of a1C transcripts generated in human cardiac and vascular cells and tissues in response to age, drugs, hormonal and pathological stimuli. We will examine whether alterations in the molecular properties of the a1C channels occur with age and as a result of cardiovascular diseases, including atherosclerosis, hypertension, cardiomyopathy, ischemia, arrhythmias and heart failure. (4) The wavelet transform analysis will be developed and applied to investigate phosphorylation and other signaling microdomains associated with the activity of human L-type Ca2+ channels. The established links will be tested with transgenic animal models. Results of our study give insights into the fundamental principles of Ca2+ signaling supporting transcription regulation as well as excitation-contraction coupling in human cardiac and vascular muscle cells and provide useful clues for molecular diagnostics and drug developments.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Intramural Research (Z01)
Project #
1Z01AG000294-07
Application #
7325058
Study Section
(LCI)
Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
2006
Total Cost
Indirect Cost
Name
Aging
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Kobrinsky, Evgeny; Soldatov, Nikolai M (2016) Electrophysiological evidences of interaction between calcium channels and PA of anthrax. Channels (Austin) 10:253-5
Soldatov, Nikolai M (2015) CACNB2: An Emerging Pharmacological Target for Hypertension, Heart Failure, Arrhythmia and Mental Disorders. Curr Mol Pharmacol 8:32-42
Kobrinsky, Evgeny; Lee, Jung-Ha; Soldatov, Nikolai M (2012) Selective fluorophore-assisted light inactivation of voltage-gated calcium channels. Channels (Austin) 6:154-6
Asemu, Girma; Fishbein, Kenneth; Lao, Qi Zong et al. (2011) Cardiac phenotype induced by a dysfunctional ? 1C transgene: a general problem for the transgenic approach. Channels (Austin) 5:138-47
Lao, Qi Zong; Kobrinsky, Evgeny; Liu, Zhuo et al. (2010) Oligomerization of Cavbeta subunits is an essential correlate of Ca2+ channel activity. FASEB J 24:5013-23
Gronich, Naomi; Kumar, Azad; Zhang, Yuwei et al. (2010) Molecular remodeling of ion channels, exchangers and pumps in atrial and ventricular myocytes in ischemic cardiomyopathy. Channels (Austin) 4:101-7
Sorond, Farzaneh A; Serrador, Jorge M; Jones, Richard N et al. (2009) The sit-to-stand technique for the measurement of dynamic cerebral autoregulation. Ultrasound Med Biol 35:21-9
Sorond, Farzaneh A; Shaffer, Michele L; Kung, Andrew L et al. (2009) Desferroxamine infusion increases cerebral blood flow: a potential association with hypoxia-inducible factor-1. Clin Sci (Lond) 116:771-9
Rudolph, James L; Sorond, Farzaneh A; Pochay, Val E et al. (2009) Cerebral hemodynamics during coronary artery bypass graft surgery: the effect of carotid stenosis. Ultrasound Med Biol 35:1235-41
Kobrinsky, Evgeny; Abrahimi, Parwiz; Duong, Son Q et al. (2009) Effect of Ca(v)beta subunits on structural organization of Ca(v)1.2 calcium channels. PLoS One 4:e5587

Showing the most recent 10 out of 32 publications