Overview of the Center for Membrane Protein Research

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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

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• 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.

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• 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
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• 
  Membership
• Guillermo Altenberg, M.D., Ph.D., Associate
  ProfessorStructure, function and regulation of
  normal gap-junctional proteins and mutants that
  cause heart disease and deafness.[phone],
  G.Altenberg_at_ttuhsc.edu Pablo Artigas, Ph.D.,
  Assistant ProfessorStructure based functional
  studies of Na/K ATPase and bilayer regulation of
  membrane protein function.[phone],
  Pablo.Artigas_at_ttuhsc.edu Michael P. Blanton,
  Ph.D., Associate ProfessorStructural analysis of
  ligand gated channels.[phone],
  Michael.Blanton_at_ttuhsc.edu Joe A. Fralick,
  Ph.D., ProfessorTransport physiology of
  bacteria.[phone], Joe.Fralick_at_ttuhsc.edu 
  Lan Guan, M.D., Ph.D., Assistant
  ProfessorHigh-resolution structure modeling of
  solute transporters.[phone],
  Lan.Guan_at_ttuhsc.edu Juyang Huang, Ph.D.,
  Associate ProfessorThe role of cholesterol in
  determining the physical, chemical and functional
  properties of biomembranes.[phone],
  Juyang.Huang_at_ttu.edu
•  

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• Michaela Jansen, Ph.D., Assistant Professor
  Structure and function studies of ligand-gated
  ion channels and transporters.[phone],
  Michaela.Jansen_at_ttuhsc.edu Jose Perez-Zoghbi,
  Ph.D., Assistant ProfessorThe cellular
  mechanisms of epithelium-smooth muscle
  communication in the lung.[phone],
  Jf.Perez_at_ttuhsc.edu Thomas A. Pressley, Ph.D.,
  ProfessorFunction and regulation of the
  sodium-potassium pump and similar ion
  transporters.[phone], Thomas.Pressley_at_ttuh
  sc.edu Luis Reuss, M.D., Professor and
  DirectorIon and water transport mechanisms
  structure and function of gap-junction channels
  and hemichannels.[phone],
  Luis.Reuss_at_ttuhsc.edu R. Bryan Sutton, Ph.D.,
  Associate Professor X-ray crystallography of
  peripheral membrane associating C2 domains in
  synaptotagmin and human dysferlin.(806)
  743-4058, Roger.B.Sutton_at_ttuhsc.edu Ina
  Urbatsch, Ph.D., Assistant ProfessorStructure-fun
  ction relationships in the multidrug-resistance
  proteins.[phone], Ina.Urbatsch_at_ttuhsc.edu

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• Joachim Weber, Ph.D., Assistant
  ProfessorEnzymatic mechanism of ATP synthesis by
  the ATP synthase.[phone],
  Joachim.Weber_at_ttuhsc.edu
• ..soon to be added
• Luis Cuello, Ph.D., Assistant ProfessorStructural
  Biology and Biophysics of Ion Channels in
  Excitable Cells.[phone],
  Luis.Cuello_at_ttuhsc.edu

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Structure Function Studies of

• Cys Loop Receptors

Proton Coupled Folate Transporter

• GABAA, GABAr, Glycine, nACh, 5-HT3

Diseases Hereditary Folate Malabsorbtioin

• Diseases Epilepsy, Anxiety, Sleep disorders, Eye
  diseases, Learning, Memory, Alzheimer, Dementia,
  Nausea during Chemotherapy

Michaela Jansen, Cell Physiology and Molecular
Biophysics
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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
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Sutton Lab

• X-ray crystallography of C2 domain proteins
• Peripheral membrane proteins
• Synaptotagmin
• present at the pre-synaptic terminal
• Ca2 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

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Piecemeal Dissection of the dysferlin protein
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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.
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Joachim Weber TTU 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. Webers 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. Webers 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.
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Isoform Diversity in the Na,K-pump
Thomas A. Pressley Department of Cell Physiology
and Molecular Biophysics
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Enzyme heterogeneity Lessons from the
Na,K-ATPase

• Presence of multiple isoforms must confer
  selective advantage, but functional relevance is
  unknown

Approaches
Enzymology
Fluorescent Labeling
Molecular Biology
Immunodetection
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Michael P. Blanton, Ph.D. Professor/ GSBS
Associate Dean Department of Pharmacology and
Neuroscience

• ONGOING RESEARCH PROJECTS AND
  COLLABORATIONS IN THE BLANTON LAB.
• Structure/Function Studies of Cys-Loop Ligand
  Gated Ion Channels (LGICs).
• Structure/Function Studies of Proton Coupled
  Folic Acid Transporter (PCFT). Blanton/Jansen
  labs.
• 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.
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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 50th
Annual Meeting of Biophysical Society
Shouryadeep
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Fall 2010 AdmissionsM.S. and Ph.D.

• Texas Tech University HSC
• -Undeclared Track(choose track at end
  of 1st 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/academ
  ics/admissions.aspx

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Fall 2010 AdmissionsM.S. and Ph.D.

• Texas Tech University
• Apply online http//www.depts.ttu.edu/gradscho
  ol/

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Questions ?