NativeMS

A two-day short course in Native Mass Spectrometry will be presented on June 1-2, 2024 in Anaheim, CA, in conjunction with the annual American Society for Mass Spectrometry Conference.  Native MS is making significant contributions to structural biology and because of its ease of application and broad applicability often leads to an integrated approach with other structural biology tools. The course is designed to introduce the broad field of native mass spectrometry to new practitioners of native MS. 

Instructors are Vicki Wysocki, David Russell, Brandon Ruotolo, Sophie Harvey, Art Laganowski, and Michael Marty. Topics to be covered include: 

• sample preparation for native MS 
• instrumentation used for native MS - how does it differ from other MS 
• spray conditions to keep soluble complexes native 
• spray conditions for membrane protein complexes (micelles, liposomes, nanodiscs) 
• activation methods for native MS (CID, CIU, SID, UVPD, ETD/ECD/EID) 
• complementary methods (HDX, covalent labeling, crosslinking, top-down MS) 
• ion mobility for native MS • online separations approaches coupled to native MS (IEX, SEC, CE) 
• software tools for native MS • (Unidec, Impact, Pulsar, EMnIM, DynamXL, XLinkX, iFAMS) 
• case studies for protein:protein, protein ligand, RNA: protein, DNA:protein, and membrane protein complexes

For more information and registration, please visit the ASMS website. 

The Native MS Guided Structural Biology Center will present a Thermo-sponsored seminar on January 16, 2024, to highlight the complementarity of native mass spectrometry and cryoEM. In this seminar, you 'll learn the basics of mass spectrometry and cryo-EM techniques and how the integration of mass spectrometry and cryo-EM can support your research. 

Bound Together: Mass Spectrometry and Cryo-EM  

January 16, 2024 | 09:30 am – 3:40 pm EST | 115 Biomedical Research Tower 

Agenda

9:30 am to 10:00 am: Registration and light breakfast (provided by Thermo Fisher) 

10:00 am to 10:45 am: Natalia de Val, Ph.D., Thermo Fisher Scientific, “Introduction to Cryo-EM and its Integration with Mass Spectrometry, Application Results” 

10:45 am to 11:30 am: Weijing Liu, Ph.D., Thermo Fisher Scientific, “Introduction to Mass Spectrometry and its Integration with Cryo-EM, Application Results” 

11:30 am to 12:15 pm: Brandon Ruotolo, Ph.D., University of Michigan, “High-throughput Collision Induced Unfolding as a Screening Technology for High-Resolution Structural Biology” 

12:15 pm to 1:15 pm: Lunch (provided by Thermo Fisher) 

1:15 pm to 2:00 pm: Chuck Bell, Ph.D., The Ohio State University, “Structure and Assembly of a Novel DNA Repair Complex by Cryo-EM and Native Mass Spectrometry” 

2:00 to 2:45 pm: Vicki Wysocki, Ph.D., The Ohio State University, “Native MS of Adeno Associated Viruses and Glycoproteins” 

2:45 pm to 3:30 pm: Michael Cianfrocco, Ph.D., University of Michigan, ”Artificial intelligence algorithms to automate cryo-EM data collection” 

3:30 pm-3:40 pm: Closing remarks

We have received notice of award of a 6.52M RM1 grant for our next five year period, converting the Biomedical Technology Research Resource (BTRR) to a national Biomedical Technology Optimization and Dissemination (BTOD) Center. The overarching goal of the Native MS Guided Structural Biology Center is to translate an integrated native mass spectrometry (nMS) workflow for characterization of macromolecular protein and nucleoprotein complexes to the biomedical research community. The tools provided for expert and non-expert users will allow them to integrate nMS approaches in synergy with other structural biology tools, taking advantage of nMS from project beginning to end, with dissemination through vendor partnerships and training. The Center works with investigators across the nation and globe on challenging biomedical projects ranging from HIV to anti-coronavirus immunogens and neurological disorders. 

The U54 Behavior of HIV in Viral Environments (B-HIVE) Center furthers the understanding of HIV-1 and its interactions with cellular host factors within distinct cellular environments that shape the HIV replication cycle. The P41 Resource has been working with the U54 team on characterizing the interactions between the HIV capsid protein and small molecules. The B-HIVE center invited collaborator Dr. Sophie Harvey to give a nMS training webinar (May 2023), covering the basics of nMS, high-resolution nMS and ion mobility. B-HIVE Center

A Workshop in Native Mass Spectrometry will be held on July 23, 2023 in conjunction with AMS 2023 to be held in Austin, Texas. Instructors are Vicki Wysocki (OSU), Dave Russell (Texas A&M University), Art Laganowsky (Texas A&M), Brandon Ruotolo (University of Michigan). 

Introduction:

Native mass spectrometry is an exploding area in the mass spectrometry community. Native MS is making significant contributions to structural biology and because of its ease of application and broad applicability, can lead and/or guide an integrated approach that also uses other structural biology tools. This course is designed to introduce the broad field to new practitioners of native MS.

Topics to be covered include:

• intro and case studies for protein:protein, protein ligand, RNA: protein, DNA:protein, and membrane protein complexes

• sample preparation for native MS

• instrumentation used for native MS -  how does it differ from other MS, activation methods

• spray conditions to keep soluble complexes native

• spray conditions for membrane protein complexes (micelles, liposomes, nanodiscs)

• ion mobility for native MS

Prerequisites: Basic knowledge of mass spectrometry, including some familiarity with ESI and mass analyzers

Vicki Wysocki was invited to present in a symposium encompassing recent advances in native MS from both academia and biopharma. The online symposium, Native Mass Spectrometry (nMS) for Studying Biomolecules, was hosted by Pharmaceutical & Bioscience Society International (PBSS), and co-organized by Dr. Surinder Kaur at Genentech and Dr. Daojing Wang at Newomics. The symposium was held March 14, 2023, with 318 participants. 

Speakers and Topics included:
Surinder Kaur (Co-organizer), PhD, Executive Director, BioAnalytical Sciences, Genentech, Welcome Remarks

Daojing Wang (Co-organizer), PhD, Founder & CEO, Newomics, Inc., Symposium Overview

Joseph A. Loo, PhD, Professor, UCLA, “Linking Native Mass Spectrometry and Top-down MS for Characterizing Proteins and Complexes”

Shunhai Wang, PhD, Director, Analytical Chemistry Group, Regeneron, “Empowering Native MS Characterization of Biologics by a Diverse Collection of Separation Techniques”

Elizabeth Hecht, PhD, Principal Scientist, Dept. of Microchemistry, Proteomics, and Lipidomics, Genentech, “Hybrid MS, IM, and Imaging Approaches to Separate, Automate, and Characterize Protein Complexes”

Rosa Viner, PhD, Sr. Manager Omics Marketing, Thermo Fisher Scientific, “Automated High-throughput Online Native MS Screening for Proteins and Protein Complexes”

Vicki Wysocki, PhD, Professor and Ohio Eminent Scholar, Ohio State University, “Electron Capture and Surface Collisions for Protein and Nucleoprotein Structure Characterization”

Vicki Wysocki will host a webinar in Native MS for Structural Biology presented by the Protein Society on March 15, 2023, featuring research talks by several leaders in the field.

Please join us March 15, 2023, from 12:00 PM-2:00 EDT / 4 -6 PM England GMT / 5- 7 PM The Netherlands CET, as we hold our 19th virtual workshop titled: Native Mass Spectrometry for Structural Biology.

Research Talks:
Vicki Wysocki, The Ohio State University – Organizer

Dame Carol Vivien Robinson, University of Oxford
Dr. Albert Heck, Utrecht University
Dr. Stephan Rauschenbach, University of Oxford  
Dr. Vicki Wysocki, The Ohio State University

Organizer and Moderator: Dr. Vicki Wysocki, The Ohio State University


Protein Society webinars are free of charge.
Space is limited. Protein Society Members have priority registration.

Time

Mar 15, 2023 12:00 PM in Eastern Time (US and Canada)

Link to register

A two-day short course will be presented on June 3-4, 2023 in Houston, TX, in conjunction with the annual American Society for Mass Spectrometry Conference. The course is designed to introduce the broad field of native mass spectrometry to new practitioners of native MS. Instructors are David Russell, Arthur Laganowsky, Brandon Ruotolo and Sophie Harvey. Topics include: Sample preparation for native MS; Membrane protein native MS; Instrumentation for nMS; Instrument conditions to keep soluble complexes native; Online separations approaches couples to nMS; Activation methods for nMS (non-covalent); Native-omics; Ion mobility instrumentation for nMS; Theoretical calculations and software for CCS determination; Intro to software tools and examples; Software tools demonstration; Variable temperature ESI; Thermodynamics and binding constants; Integrated platform and case studies. Please visit the ASMS website for more information.

Wysocki Group

• Chen Du, Samantha H. Sarni, Jorjethe Roca, Mengxuan Jia, Ewelina Malecka-Grajek, Maggie Li, Sarah Woodson, Vicki H. Wysocki, Native MS and SID reveal that RNA binding lowers the energy required for dissociation of the RNA chaperone HFQ, 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN (oral)
• Sean Cleary, Chen Du, Marius M Kostelic, Michael T. Marty, Nicholas Horvath, Vicki H. Wysocki, Adeno-Associated Virus Characterization by SID-CDMS, 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN (oral)
• Ila Marathe, Stella Lai, Venkat Gopalan, Vicki Wysocki. Use of native mass spectrometry to establish a robust calibration curve for mass photometry-based measurements of RNAs and ribonucleoprotein complexes, 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN (oral)
• Stella M. Lai, Ila A. Marathe, Venkat Gopalan, and Vicki H. Wysocki, Minimizing 3′-heterogeneity in in vitro transcribed RNAs for native mass spectrometry-based analyses of RNA and RNA–protein complexes, 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN (oral)
• Leon (Yu-Fu) Lin, Yuan Gao, Arpad Somogyi, Vicki Wysocki; Do Different Surface Materials Influence the Surface Collision Energy Required for Dissociation and Restructuring of Protein Complexes?, ASMS Minneapolis, June 2022 (poster)

Russell Group

• Sumeet S. Chakravorty; Robert L. Schrader; Thomas E. Walker; David H. Russell; Peter T. A. Reilly. “Experimentally Adding DC to Digital Mass Filters in Higher Stability Zones.” 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN.
• Robert L. Schrader; Thomas E. Walker; Sumeet Chakravorty; Peter T. A. Reilly; David H. Russell. “Development of Charge Detection Mass Spectrometry on a Native Digital Waveform Dual-Quadrupole FT-IMS-Orbitrap Mass Spectrometer.” 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN
• Liqi Fan; Thomas E Walker; He Sun; Syuan-Ting Kuo; Robert L. Schrader; David H. Russell.“Structure and dynamics of transthyretin, a variable-temperature ESI study on the effects of mutants on stability and subunit exchange.” 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN
• Thomas E Walker; He Sun; Syuan-Ting Kuo; Liqi Fan; Robert L. Schrader; David H. Russell. ”Determination of Entropy and Enthalpy in the Binding of ATP by GroEL.” 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN
• Thomas E Walker; He Sun; Syuan-Ting Kuo; Liqi Fan; Robert L. Schrader; David H. Russell (oral). “The role of entropy in TMAO-induced stabilization of peptides.” 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN
• Thomas E Walker; Robert L. Schrader; Nate Buzitis; Gordon Anderson; Brian H. Clowers; David H. Russell; Jinyuan Liu. “An All-in-One Power Supply for the Operation of Low-Pressure Drift Tube Ion Mobility-Mass Spectrometry Systems.” 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN
• He M Sun; Thomas E Walker; Syuan-Ting Kuo; Liqi Fan; Robert L. Schrader; David H. Russell. “Investigation of GroEL-ES Complexes Thermal Stability using Native Mass Spectrometry.” 70th ASMS Conference on Mass Spectrometry and Allied Topics, June 5 - 9, 2022, Minneapolis, MN

https://www.asms.org/conferences/annual-conference/online-planner-app

Vicki Wysocki will present a one hour webinar, “Automating Native Mass Spectrometry Through the Use of Online Buffer Exchange” on March 23, 2022, sponsored by Thermo Fisher Scientific.   

Native mass spectrometry (nMS), in which intact protein or nucleoprotein complexes are directly measured, commonly involves exchange of nonvolatile electrolytes (NaCl, PBS, Tris) for volatile electrolytes such as ammonium acetate. This can be time-consuming if offline buffer exchange spin cartridges are used.  Furthermore, some samples may not be stable if left in nonvolatile buffer for more than just a very short period of time. This problem can be avoided if an LC-compatible online buffer exchange column is used to separate protein complex from salt.  In this experiment, no attempt is made to perform size exclusion chromatography to separate protein complexes of different sizes but rather any protein complex that is injected onto the system is simply separated from low mass salts that would otherwise product heavily adducted protein complexes that appear as broad peaks in the mass spectrum.  This webinar will focus on the development of online buffer exchange (OBE) for nMS applications and will describe how the experiment is implemented and how it can be extended by coupling to affinity separation (e.g., IMAC-OBE) to be used to optimize protein overexpression.  

Key Learning Objectives: 

• The use of online buffer exchange for high throughput native mass spectrometry   

• Online buffer exchange nMS allows to directly screen structural features of large biomolecules even if such biomolecules are not very stable in MS friendly buffers  

• How the coupling of affinity separation (e.g., IMAC-OBE) to online-buffer exchange can be used to optimize protein overexpression  

Who Should Attend: 

• MS practitioners  

• Researchers/ R&D Managers  

• Laboratory Managers/ Directors / Supervisors  

• Laboratory Technicians / Operators  

• Structural Biologists  

• Scientists in biopharma engaged in biotherapeutic characterization 

A two-day short course in Native MS will be presented at ASMS 2022 in Minneapolis, MN on June 4-5, 2022.

The Wysocki group will attend the 69th ASMS Conference on Mass Spectrometry and Allied Topics in Philadelphia, PA October 31 – November 4, 2021.

Please stop by and see our oral and poster presentations if you are attending ASMS in person!

• Oral: Implementation of surface-induced dissociation on a digital quadrupole selection system coupled to an Orbitrap EMR via the HCD cell given by Benjamin Jones in session MOF pm at 02:30 pm on Mon, Nov 01
• Oral: Resolving Macromolecular Complexes using Surface Induced Disassociation and Charge Detection Native Mass Spectrometry given by Sean Cleary in session TOF am at 09:10 am on Tue, Nov 02
• Oral: Fundamental aspects of protein complex activation by SID: stopping curves, ERMS, and HDX given by Dalton Snyder in session WOF am at 08:50 am on Wed, Nov 03
• Oral: Post-translational modifications induced by Salmonella infection with multi PTM enrichment workflow and open search bottom-up proteomics given by Yongseok Kim in session WOB pm at 03:10 am on Wed, Nov 03
• Poster (MP 128): Leveraging tandem affinity-buffer exchange chromatography online with native mass spectrometry to characterize overexpression cell lysates and purified proteins/protein complexes given by Stella Lai on Mon, Nov 01
• Poster (MP 302): Probing the structure and interactions of membrane protein complexes using native mass spectrometry and surface induced dissociation given by Sophie Harvey on Mon, Nov 01
• Poster (TP 314): Proteomics Characterization of Mouse Feces During Salmonella Infection given by Maryam Baniasad on Tue, Nov 02
• Poster (WP 182): TIMS-SID-TIMS for structural determination of protein assemblies given by Yu-Fu Lin on Wed, Nov 03
• Poster (ThP 284): Native mass spectrometry and surface-induced dissociation reveal connectivity of the disordered C-terminal domain with the structured Sm core of Hfq give by Chen Du on Thu, Nov 04

A two-day short course in Native MS will be presented at ASMS 2021 on October 30-31, 2021.

Vicki Wysocki, Sophie Harvey, David Russell, and Arthur Laganowsky will serve as instructors. Topics include: Sample preparation for native MS; Membrane protein native MS; Instrumentation for nMS; Instrument conditions to keep soluble complexes native; Online separations approaches couples to nMS; Activation methods for nMS (non-covalent); Native-omics; Ion mobility instrumentation for nMS; Theoretical calculations and software for CCS determination; Intro to software tools and examples; Software tools demonstration; Variable temperature ESI; Thermodynamics and binding constants; Integrated platform and case studies. 

Supplement grant from NIH to fund purchase of a Refeyn TwoMP Mass Photometer

A notice of award was received from the NIH for a Supplement grant to the P41 Resource for Native MS-guided Structural Biology for purchase of a Refeyn TwoMP Mass Photometer. Ohio State's College of Arts and Sciences and ERIK each contributed funds. The benchtop Refeyn mass photometer will complement existing structural biology equipment at Ohio State by offering interference spectrometry measurements of protein complexes. The mass photometry equipment will initially be housed in the Wysocki lab for use in the P41 program by its Driving Biomedical Projects and Collaboration and Service projects. 

The Russell group leads development of a new variable-temperature ESI device for solution-phase studies of cold- and heat-induced folding, denaturation and self-assembly of proteins and protein complexes.

Recently the P41 Team has developed a new variable-temperature ESI device for solution-phase studies of cold- and heat-induced folding, denaturation and self-assembly of proteins and protein complexes. Using this device, the sample solution contained within the ESI emitter can be varied from 275K to 373K, which allows for direct sample of the effects of the solution environment (Gibbs free energy) on the structure/conformational preferences of the sample. The capabilities of the vT-ESI source have been demonstrated using model proteins (ubiquitin and human frataxin) and large protein complexes (GroEL, GroES complexes and the effects of ligands on the structure and stability of the intact GroEL/GroES complex. The vT-ESI source design and materials have been disseminated to a number of research groups and collaborators, including Arthur Laganowsky (Texas A&M University), Vicki Wysocki (The Ohio State University), David Clemmer (University of Indiana), Joseph S. Beckman (Oregon State University and eMSions), and Jennifer Brodbelt (University of Texas).

Advances in software tools for native MS will be presented by software developers on June 8-10, 2021.

The Resource will present a three-day workshop on June 8-10, 2021 (virtual). Software tool developers will present their advances in software tools with applications for native MS, top-down, collision cross sections, ion mobility, cross-linking and more. The workshop will include speaker presentations and breakout sessions for demonstrations and office hours. Presenters and session chairs include:

• Michael Marty/Marius Kostelic, University of Arizona (UniDec)
• Ken Durbin/Jack McGee (Kelleher group), Northwestern University (ProSight Native)
• Christian Bleiholder, Florida State Univeristy (PSA)
• Brandon Ruotolo, University of Michigan (CIUSuite)
• Michal Sharon, Weizmann Institute (session chair)
• Marshall Bern/Grace Zdeblick, Protein Metrics (Intact Mass)
• Oliver Kohlbacher/Kyowon Jeong, Universität Tübingen (FlashDeconv)
• Erik Marklund, Uppsala Universitet (IMPACT)
• Perdita Barran, University of Manchester (session chair)
• Jim Prell, University of Oregon (iFAMS, Collidoscope)
• Sean McIlwain, University of Wisconsin-Madison (MASH Explorer)
• Steffen Lindert/Bargeen Turzo, The Ohio State University (MS-guided structural prediction with Rosetta)
• Matteo Degiacomi, Durham Univerisity (EM∩IM , DynamXL)
• Ying Ge, University of Wisconsin-Madison (session chair)

The Wysocki Group and the Russell Group are looking for well-qualified postdoctoral researchers to help advance the missions of the Resource for Native Mass Spectrometry Guided Structural Biology.  If you are interested in learning more about postdoctoral training and/or other training opportunities, please submit a training request.

A three hour workshop, Introduction to Native Mass Spectrometry, will be presented on October 1, 2019 in advance of the 3^rd annual Ohio Mass Spectrometry and Metabolomics Symposium, held at The Ohio State University in Columbus, Ohio. P41 Resource personnel Florian Busch, Dalton Snyder, Sophie Harvey, Zac VanAernum, Alyssa Stiving and Vicki Wysocki will present an introduction to native MS methods used to help solve structural biology questions. Workshop topics include preparation/online separations, spray and instrument conditions, ion mobility, activation methods, and case studies. 

Resource to present preconference Native MS Workshop at AMS in Amherst, MA on July 21, 2019

In a first-of-its kind scientific breakthrough, scientists have programmed proteins to pair exactly, similar to the way DNA molecules zip together.

A $6.8 million NIH grant has established a Resource for Native MS-Guided Structural Biology at Ohio State under the direction of Vicki Wysocki.