X-Ray Footprinting at the ALS: New Opportunities in Structural Biology

X-ray Footprinting as a Stand Alone and Component of Hybrid Techniques to Resolve Structure and Dynamics of Proteins in Solutions

Organizers: Jen Bohon, Sayan Gupta, Corie Ralston

ALS Users’ Meeting 2014
Date: Oct 8, 2014
Venue: Lawrence Berkeley National Laboratory, Building 6, room 1105

Workshop Description

X-Ray Footprinting (radiolytic hydroxyl radical labeling and mass spectrometry) is a technique that provides structural and dynamical information on proteins in the solution state and is highly complementary to many other structural and biochemical techniques. Because it yields residue-specific information by in situ generation of hydroxyl radicals, it is used to determine conformation changes, ligand binding and protein-protein interactions under various conditions at a high degree of both temporal and spatial resolution and therefore is useful to both fundamental research and characterization of biologics/protein therapeutics. The technique was pioneered at the NSLS a decade ago and is now available at the ALS with advanced performance. This workshop will cover the basics and recent advances of the x-ray footprinting experiment: how to prepare samples, how to analyze data, and the information that can be determined through the experiment. In addition, several talks will cover how to combine XFP with other more well-known structural techniques, such as macromolecular crystallography, SAXS, NMR, and cryo-EM. The workshop will be useful for biological researchers in academia and the pharmaceutical industry.

Agenda

TIME	SESSION
9:30am-11:45am	Methods and Tutorials
	Welcome and Overview of the XFP Method – Link to Talk Corie Ralston, Lawrence Berkeley National Laboratory XF Mass Spectrometry and Advances in Data Analysis Approaches – Link to Talk Janna Kiselar, Case Western Reserve University The Hybrid Approach: Combining XFP with SAXS and MX – Link to Talk Sichun Yang, Case Western Reserve University ProtMapMS: Software Solutions for High-Throughput Examination of Covalently Labeled Biomolecules by Structural Mass Spectrometry – Link to Talk Parminder Kaur, Case Western Reserve University Coffee and Discussion Everyone!
11:45am-1:00pm	Lunch
1:00pm-3:00pm	Recent Exciting Results
	Probing Ribosome Assembly in Live Cells Sarah Woodson, Johns Hopkins University A Synchrotron-based Hydroxyl Radical Footprinting Analysis of Amyloid Fibrils and Prefibrillar Intermediates with Residue-specific Resolution Janna Kiselar, Case Western Reserve University Using XRF to Probe Protein Conformational Changes Governing Photoprotection in Cyanobacteria – Link to Talk Cheryl Kerfeld, Michigan State University Visualizing Internal Water Interactions in Membrane Proteins by X-ray Footprinting – Link to Talk Sayan Gupta, LBNL Coffee and Discussion Everyone!
3:00pm-4:00pm	More Exciting Results: Antibodies and HIV
	XFP Insights into IgG Galahad Deperalta, Genentech Unmasking the initial stages of HIV Env glycoprotein activation using H/D exchange and X-ray footprinting – Link to Talk Miklos Guttman, University of Washington, Seattle
4:00pm-5:00pm	The Future is Here
	The NSLS-II XFP Beamline and Beyond – Link to Talk Jen Bohon, Case Western Reserve University/Brookhaven National Laboratory Tour of Beamlines and Discussion Jen Bohon, Corie Ralston, Sayan Gupta