Transcript Overview of the Center for Membrane Protein Research Michael P. Blanton -
Overview of the Center for Membrane Protein Research
Presented by
Michael P. Blanton
Associate Professor in the Department of Pharmacology and Neuroscience -Associate Dean of the Graduate School of Biomedical Sciences (GSBS) -Member of the CMPR Steering Committee
• The Center for Membrane Protein Research (CMPR) established in 2007, is one of 11 Centers of Excellence in the School Medicine (SOM) at Texas Tech University Health Sciences Center (TTUHSC).
• Dr. Luis Reuss is the Director of the CMPR • Currently the CPMR has 13 faculty members that come from four different basic science departments within TTUHSC and 2 departments within TTU.
Mission
• The long-term goal of the Center is to advance our knowledge of the structure and function of membrane proteins in health and disease. The Center brings together a group of TTUHSC and TTU investigators interested in the broad field of membrane-protein research.
•
Rationale:
After completion of the human genome sequence, biomedical research has evolved into a combination of genomics, proteomics, and functional genomics. To a great extent, biomedical research in this century will be focused on prototypical proteins and protein families, including the determination of their structures, normal function, and their roles in human disease. From this knowledge will emanate rational design of new pharmacological agents that will open novel therapeutic approaches.
About 30% of the genes included in the human genome encode membrane proteins. These proteins participate in a myriad of normal and abnormal cell functions, including: 1) transport of ions, water and small solutes; 2) signaling processes; 3) metabolism and detoxification; 4) programmed cell death and necrosis; 5) entry of pathogens into cells, and 6) cellular structural integrity. If the promise of modern proteomics is to be fully realized, greater attention must be paid to the structures of these proteins and how they relate to normal and abnormal function. Crystallization is the method of choice for generating high-resolution structural models. However, membrane proteins have both hydrophobic and hydrophilic surfaces, a duality that makes them more difficult to crystallize than water-soluble proteins. It follows that relatively few structures of membrane proteins have been solved at the level of atomic resolution. In addition, high-resolution structures are important but not sufficient to understand how membrane proteins (and soluble proteins as well) function. To assess function, it is necessary to carry out biochemical and biophysical studies that are informed by structural knowledge, but explore questions of molecular mechanism, protein-protein interactions, and regulation.
http://www.ttuhsc.edu/som/physiology/programs/cmpr.aspx
• •
Membership Guillermo Altenberg, M.D., Ph.D.
(806) 743-2531,
, Associate Professor
Structure, function and regulation of normal gap-junctional proteins and mutants that cause heart disease and deafness.
Pablo Artigas, Ph.D.
(806) 743-3170,
, Assistant Professor
Structure based functional studies of Na/K ATPase and bilayer regulation of membrane protein function.
Michael P. Blanton, Ph.D.
(806) 743-2526,
, Associate Professor
Structural analysis of ligand gated channels.
Joe A. Fralick, Ph.D.
(806) 743-2555,
, Professor
Transport physiology of bacteria.
Lan Guan, M.D., Ph.D.
(806) 743-2520,
, Assistant Professor
High-resolution structure modeling of solute transporters.
Juyang Huang, Ph.D.
(806) 742-4780,
, Associate Professor
The role of cholesterol in determining the physical, chemical and functional properties of biomembranes.
•
Michaela Jansen, Ph.D.
transporters.
(806) 743-2520,
, Assistant Professor
Structure and function studies of ligand-gated ion channels and [email protected]
Jose Perez-Zoghbi, Ph.D.
the lung.
(806) 743-2522,
, Assistant Professor
The cellular mechanisms of epithelium-smooth muscle communication in [email protected]
Thomas A. Pressley, Ph.D.
transporters.
(806) 743-4056,
, Professor
Function and regulation of the sodium-potassium pump and similar ion [email protected]
Luis Reuss, M.D.
(806) 743-2627,
, Professor and Director
Ion and water transport mechanisms; structure and function of gap-junction channels and hemichannels.
R. Bryan Sutton, Ph.D., Associate Professor
X-ray crystallography of peripheral membrane associating C2 domains in synaptotagmin and human dysferlin.
(806) 743-4058, [email protected]
Ina Urbatsch, Ph.D.
(806) 743-2700,
, Assistant Professor
Structure-function relationships in the multidrug-resistance proteins.
•
Joachim Weber, Ph.D.
, Assistant Professor
Enzymatic mechanism of ATP synthesis by the ATP synthase.
(806) 742-1297, [email protected]
• ……………..soon to be added:
Luis Cuello, Ph.D.
, Assistant Professor
Structural Biology and Biophysics of Ion Channels in Excitable Cells.
(806) 742-1297, [email protected]
Structure Function Studies of
•
Cys Loop Receptors GABA
A
, GABA
r
, Glycine, nACh, 5-HT
3
Proton Coupled Folate Transporter
Diseases: Epilepsy, Anxiety, Sleep disorders, Eye diseases, Learning, Memory, Alzheimer, Dementia, Nausea during Chemotherapy Diseases: Hereditary Folate Malabsorbtioin Michaela Jansen, 5A172
Michaela Jansen, Cell Physiology and Molecular Biophysics
• • • • • • • • •
Questions Asked / Answered
Which part of the protein lines the solute pathway?
Which amino acids are involved in ligand binding? How much do certain protein parts move in the resting state? How do they move during gating?
How is binding of ligand in the binding site transduced to opening of the gate?
How do disease causing mutations affect function?
Techniques Utilized
Molecular Biology Heterologous Expression / bacteria, mammalian cells, Xenopus oocytes Biochemistry / Western Blotting Electrophysiology
Michaela Jansen, Cell Physiology and Molecular Biophysics
Michaela Jansen, 5A172
X-ray crystallography of C2 domain proteins
Peripheral membrane proteins
Synaptotagmin
present at the pre-synaptic terminal Ca +2 sensor for exocytosis
Human dysferlin
Limb-Girdle Muscular Dystrophy Caused by mutations within the dysferlin gene Patients are typically wheelchair-bound by 30 years of age
N
A B -Type 2 -Type 1 C D -Putative hexamerization domain E F G TM
C
Guillermo Altenberg, M.D., Ph.D.
Associate Professor Cell Physiology and Molecular Biophysics
My laboratory is interested on the mechanisms of transport across biological membranes, with a focus on membrane protein structure, function and regulation.
Our target proteins are connexins, the gap junction-forming proteins, and multidrug resistance proteins of the ATP-binding cassette (ABC) superfamily, which export a number of chemically dissimilar compounds from the cells.
Ongoing projects
1) Molecular mechanisms of regulation of the permeability Cx43 gap-junction channels by calcium calmodulin and PKC-mediated phosphorylation.
2) Mechanisms of deafness caused by mutations of Cx26. Mutations of Cx26 are the most frequent cause of deafness.
3) Molecular mechanism of ABC exporters studied by determining how different domains of the proteins move during the transport cycle.
We use molecular a variety biological (protein purification and reconstitution), physiological (transport assays) and biophysical (electrophysiology, fluorescence, of methodologies, (mutagenesis), including biochemical luminescence transfer) techniques.
resonance energy
TTU
Joachim Weber – Chemistry and Biochemistry
ATP synthase
is the central enzyme in the energy metabolism of most, if not all, living organisms. It is also the smallest known rotary motor. It converts electrochemical energy (transmembrane proton gradient) into mechanical energy (subunit rotation) and back into chemical energy (ATP synthesis); it can also run in reverse, hydrolyzing ATP to generate a proton gradient.
Dr. Weber’s laboratory studies the coupling between ATP hydrolysis/synthesis and subunit rotation in residue level detail. The applied techniques are a combination of biophysical chemistry (especially fluorescence spectroscopy), molecular biology, molecular modeling, and biochemical analysis.
The goal of Dr. Weber’s research is to elucidate the mechanism of ATP synthase, to improve our understanding of the cellular energy metabolism, and to facilitate the use of this enzyme as motor in nanotechnological applications Supported by NIH grant GM071462.
Isoform Diversity in the Na,K-pump
Thomas A. Pressley Department of Cell Physiology and Molecular Biophysics
Enzyme heterogeneity: Lessons from the Na,K-ATPase
•
Presence of multiple isoforms must confer selective advantage, but functional relevance is unknown Approaches: Enzymology Immunodetection Molecular Biology Fluorescent Labeling
Michael P. Blanton, Ph.D.
Professor/ GSBS Associate Dean Department of Pharmacology and Neuroscience
ONGOING RESEARCH PROJECTS AND COLLABORATIONS IN THE BLANTON LAB. 1. Structure/Function Studies of Cys-Loop Ligand Gated Ion Channels (LGICs).
2. Structure/Function Studies of Proton Coupled Folic Acid Transporter (PCFT). Blanton/Jansen labs.
3. Examining the Lipid-Protein Interface and Lipid Protein Interactions of the Gap Junctional protein Connexin 43. Blanton/Altenberg/Huang labs.
Hamouda et al. (2007)
Biochemistry 46
, 13837-13846.
The Blanton Lab Philosophy: This is a group effort!
The Grand Canyon in January!- we drove from Lubbock, TX to Long Beach, CA to attend 50 th Annual Meeting of Biophysical Society Shouryadeep
Fall 2010 Admissions M.S. and Ph.D.
• Texas Tech University HSC -Undeclared Track
(choose track at end of 1 st year)
- Declared Track
-Cell Physiology and Molecular Biophysics -Pharmacology and Neuroscience -Microbiology and Immunology -Cell and Molecular Biology - Biochemistry and Molecular Genetics - Biotechnology (M.S. only)
Apply online
http://www.ttuhsc.edu/gsbs/academics/admissions.aspx
Fall 2010 Admissions M.S. and Ph.D.
• Texas Tech University
Apply online http://www.depts.ttu.edu/gradschool/