Abstract

Adequate amounts of a variety of high quality, purified protein are common requirements of each of the projects of this proposal. The Protein Core will use its expertise in molecular biology, protein expression, and protein purification to provide recombinant proteins for use in each of the projects of this Proposal. We have successfully fulfilled this role during the current funding period. The Protein Core has two main functions: (1) Generate purified recombinant proteins as needed by each of the projects. Optimal methods for expression, purification and, if necessary, reconstitution will be developed depending on the experimental requirements for the particular protein. Emphasis will be placed on maintaining structure, activity and function as similar as possible to native proteins. Toward these ends, extensive biochemical and functional characterizations will be done. Because several of the proposed studies consider the role of post-translational modifications (for example, phosphorylation or prenylation), careful attention will be given to assure homogeneous preparations of properly modified proteins. In addition, we will generate and purify antibodies against whole proteins or peptides. (2) Provide molecular biology support, which will include construction of expression plasmids, site-directed mutagenesis, yeast 2-hybrid assays to test for interacting proteins, and isolation of cDNA clones for proteins identified or needed by the projects.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL048807-15
Application #
7589859
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Project Start
Project End
Budget Start
2008-04-01
Budget End
2009-03-31
Support Year
15
Fiscal Year
2008
Total Cost
$241,413
Indirect Cost

  Institution

Name
University of Virginia
Department
Type
DUNS #
065391526
City
Charlottesville
State
VA
Country
United States
Zip Code
22904

  Publications

Utepbergenov, Darkhan; Derewenda, Urszula; Olekhnovich, Natalya et al. (2012) Insights into the inhibition of the p90 ribosomal S6 kinase (RSK) by the flavonol glycoside SL0101 from the 1.5 Å crystal structure of the N-terminal domain of RSK2 with bound inhibitor. Biochemistry 51:6499-510
Hoofnagle, Mark H; Neppl, Ronald L; Berzin, Erica L et al. (2011) Myocardin is differentially required for the development of smooth muscle cells and cardiomyocytes. Am J Physiol Heart Circ Physiol 300:H1707-21
Jin, Li; Gan, Qiong; Zieba, Bartosz J et al. (2010) The actin associated protein palladin is important for the early smooth muscle cell differentiation. PLoS One 5:e12823
Khromov, Alexander; Choudhury, Nandini; Stevenson, Andra S et al. (2009) Phosphorylation-dependent autoinhibition of myosin light chain phosphatase accounts for Ca2+ sensitization force of smooth muscle contraction. J Biol Chem 284:21569-79
Jin, Li; Hastings, Nicole E; Blackman, Brett R et al. (2009) Mechanical properties of the extracellular matrix alter expression of smooth muscle protein LPP and its partner palladin; relationship to early atherosclerosis and vascular injury. J Muscle Res Cell Motil 30:41-55
Cierpicki, Tomasz; Bielnicki, Jakub; Zheng, Meiying et al. (2009) The solution structure and dynamics of the DH-PH module of PDZRhoGEF in isolation and in complex with nucleotide-free RhoA. Protein Sci 18:2067-79
Jin, Li; Yoshida, Tadashi; Ho, Ruoya et al. (2009) The actin-associated protein Palladin is required for development of normal contractile properties of smooth muscle cells derived from embryoid bodies. J Biol Chem 284:2121-30
Zheng, Meiying; Cierpicki, Tomasz; Momotani, Ko et al. (2009) On the mechanism of autoinhibition of the RhoA-specific nucleotide exchange factor PDZRhoGEF. BMC Struct Biol 9:36
Jelen, Filip; Lachowicz, Pawel; Apostoluk, Wlodzimierz et al. (2009) Dissecting the thermodynamics of GAP-RhoA interactions. J Struct Biol 165:10-8
Freitas, Maria Regina; Eto, Masumi; Kirkbride, Jason A et al. (2009) Y27632, a Rho-activated kinase inhibitor, normalizes dysregulation in alpha1-adrenergic receptor-induced contraction of Lyon hypertensive rat artery smooth muscle. Fundam Clin Pharmacol 23:169-78

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