Cambridge Healthtech Institute’s
A Valuable Resource and Target
January 21-22, 2015
Membrane proteins are the gateways to the cell and are valuable drug targets. For researchers to design better-targeted drugs, they need to know their structure and functional characteristics. However, determining these characteristics is difficult, because researchers need large amounts of membrane protein to characterize their functions and map their structures. Sources are scarce due to the practical problems of working with membrane proteins—specifically, difficulties in expression, purification and crystallization. Cambridge Healthtech Institute’s Membrane Proteins conference addresses the strategies and solutions for their extraction, expression and purification, and features case studies showcasing their value as an antibody drug target. Join the in-depth exploration of how to obtain functional membrane proteins, and learn more about this important protein class.
Day 1 | Day 2 | Download
Brochure | Speaker Biographies
TUESDAY, JANUARY 20
1:30 pm Conference Registration
2:00 BuzZ Session
3:00 Refreshment Break in the Exhibit Hall with Poster Awards
3:45 BuzZ Session B
(More Details >>)
4:30-5:00 Short Course Registration
5:00-8:00 Dinner Short Courses More Details >>
WEDNESDAY, JANUARY 21
7:30 am Conference Registration and Morning Coffee
8:15 Chairperson’s Opening Remarks
William Gillette, Ph.D., Senior Scientist, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research
8:20 The Distinct Issues of Membrane Proteins versus Soluble Proteins
Robert Stroud, Ph.D., Professor, Biochemistry/Biophysics and Pharmaceutical Chemistry; Director, Membrane Protein Expression Center, University of California at San Francisco (UCSF)
Membrane proteins present distinct issues versus soluble proteins; pure, homogeneous and stable in solution. Eukaryotic proteins need expression in eukaryotic cells. These include yeast, and insect cells. Human proteins, to be expressed for antibody preparation, require proper tailoring and glycosylation; and can often be achieved only in human cells. Antibody partners can be the basis for proof of principle therapeutics, and for helping to purify, assay, and validate membrane proteins’ mechanisms.
9:00 Expression of Transmembrane Proteins in Yeast for Genetic, Biochemical and Structural Studies
Mark E. Dumont, Ph.D., Professor, Biochemistry and Biophysics, University of Rochester Medical Center
The difficulty of expression, solubilization, and purification of transmembrane proteins, particularly those from eukaryotes, constitutes a significant barrier to understanding their mechanisms. The use of a system based on expression of membrane proteins from various sources in baker’s yeast has allowed the use of genetic approaches for functional characterization and stabilization of expressed proteins, while facilitating purification of transmembrane enzymes, transporters, and receptors for biochemical studies and x-ray crystallographic structure determination.
9:30 A Novel Strategy to Express and Purify Highly Stable and Functional GPCR Signaling Complexes
Arun Shukla, Ph.D., Assistant Professor, Medicine, Duke University
Structural characterization of GPCR signaling complexes remains a very challenging but extremely important task. Molecular visualization not only reveals fundamental mechanism of assembly and functioning of such complexes but also provides a unique framework for drug discovery. We present a novel strategy to assemble a highly stable and functionally competent GPCR signaling complex in-cellulo and its subsequent purification for direct structural characterization.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing
10:50 GPCR Purification: Protein Engineering and Process Optimisation are Key Elements for Successful GPCR Crystallisation
Markus Koglin, Ph.D., Associate Director, Protein Engineering, Heptares Therapeutics
The generation of Stabilised G-protein Coupled Receptors (StaRs) has opened new routes in the purification procedures for GPCRs. Here we demonstrate that protein quality is drastically improved with increasing thermostability. StaR generation alone normally does not provide suitable material for crystallisation. The combination of StaR technology, systematic construct design, and careful optimisation of the purification process delivers a powerful approach to generate protein with suitable crystallisation qualities.
11:20 Strategies for Detergent Stabilization and Large-Scale Affinity Purification of the Functional G Protein-Coupled Receptors
Alexei Yeliseev, Ph.D., Staff Scientist, Group Leader, LMBB, NIAAA, National Institutes of Health (NIH)
Human cannabinoid receptor CB2, a G protein-coupled receptor involved in regulation of immune response, is an important target for pharmaceutical drug development. For high resolution structural studies, milligram quantities of pure, homogenous and functional protein are required. We expressed the functional CB2 receptor in E. coli, and optimized its purification by tandem affinity chromatography. An efficient strategy for stabilization of the receptor in detergent micelles and reconstituted into lipid bilayers was developed.
11:50 Overcoming Difficult to Express Proteins: Cell-Free Additives for Solubilizing and Characterizing Membrane Proteins
Matthew Coleman, Ph.D., Deputy Group Leader, Molecular Toxicology, Lawrence Livermore National Laboratory; Senior Scientist, University of California at Davis
We have developed Cell-free methods for producing membrane proteins, which are inherently difficult to obtain. These include nanolipoprotein particles (NLPs), telodendrimers, which can be combined with NLPs or lipids, and amphipathic peptides. I will further discuss how we use the three different additives and their ability to support membrane protein solubility as well as function. These processes are easily adapted to high-throughput technologies for feeding the membrane structural biology pipeline.
12:20 pm Sponsored Presentation (Opportunity Available)
12:50 Session Break
1:00 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
2:00 Chairperson’s Remarks
Ian Hunt, Ph.D., Group Leader, Proteomic Chemistry and Head, Protein Sciences, Novartis
2:05 Designer Surfactant-Like Self-Assembling Peptides for Membrane Protein Purification and Stabilization
Sotirios Koutsopoulos, Ph.D., Research Scientist, Center for Biomedical Engineering, Massachusetts Institute of Technology
Membrane proteins are integral proteins of the cell membrane and are directly involved in the regulation of many biological functions and in drug targeting. However, our knowledge of membrane proteins is limited due to difficulties in producing sufficient quantities of soluble, functional and stable receptors. Designer, surfactant-like peptides may be used to extract the protein from the cell membrane and stabilize the protein outside the membrane bilayer for further studies.
2:35 Production of Human Integral Membrane Proteins in Mammalian Cells
James D. Love, Ph.D., Director, Technology Development, Biochemistry, Albert Einstein College of Medicine
Integral membrane proteins are key targets in the understanding of health and human disease, yet producing functional material in great enough quantities for structural studies remains a formidable task. This talk highlights the expression technologies under development that will greatly aid high-throughput efforts to produce functional human membrane proteins and complexes, specifically GPCRs, for a plethora of structural techniques.
3:05 Advances for Maximizing Protein Yields in HEK293 and CHO Transient Expression
Henry C. Chiou, Ph.D., Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific
Increasing focus on complex proteins in advanced research elevates the need
for higher yields from mammalian transient expression systems. To achieve
coordinated development and synergy between novel high-capacity media,
higher productivity HEK293 and CHO cells, and higher performance transfection
reagents. This presentation will demonstrate the effectiveness of this “holistic”
approach to achieve gram-level protein yields by transient expression.
3:35 The Need for Technology Platforms in USP Development
Hugo de Wit, CEO, Cellca
Are you faced with budget reductions for USP development or even anticipated to do “more with less”? Do you see increasing expectations from market and management regarding reducing timelines and cost of goods attributes? We demonstrate how a robust USP platform can help you meet these challenges.
3:50 Sponsored Presentation (Opportunity Available)
4:05 Refreshment Break
4:25 Plenary Keynote Session
From Yeast to the Brain: Advances in Proteomics (More Details >> )
John R. Yates, Ph.D., Ernest W. Hahn Professor, Chemical Physiology and Molecular and Cellular Neurobiology, The Scripps Research Institute
5:45-7:00 Reception in the Exhibit Hall with Poster Viewing
Day 1 | Day 2 | Download
Brochure | Speaker Biographies