• Skip to main content
  • Skip to after header navigation
  • Skip to site footer
ERN: Emerging Researchers National Conference in STEM

ERN: Emerging Researchers National Conference in STEM

  • About
    • About AAAS
    • About NSF
    • About the Conference
    • Project Team
    • Advisory Board
  • Conference
  • Abstracts
    • Abstract Submission Process
    • Abstract Submission Guidelines
    • Presentation Guidelines
  • Travel Awards
  • Resources
    • Award Winners
    • Code of Conduct-AAAS Meetings
    • Code of Conduct-ERN Conference
    • Conference Agenda
    • Conference Materials
    • Conference Program Books
    • ERN Photo Galleries
    • Events | Opportunities
    • Exhibitor Info
    • HBCU-UP PI/PD Meeting
    • In the News
    • NSF Harassment Policy
    • Plenary Session Videos
    • Professional Development
    • Science Careers Handbook
    • Additional Resources
    • Archives
  • Engage
    • Webinars
    • 2023 ERN Recap Video
    • ERN 10-Year Anniversary Videos
    • Plenary Session Videos
  • Contact Us
  • Login

Protein Expression and Purification Combined with FPOP to Identify GCaMP2 Structure

Graduate #38
Discipline: Chemistry and Chemical Sciences
Subcategory: Chemistry (not Biochemistry)

Cord Carter - Jackson State University
Co-Author(s): Emily Hart, Maissa Gaye, Lisa Jones, Indiana University Purdue University Indianapolis, Indianapolis, IN



The work presented here is focused on the characterization of the fusion protein, GCaMP2. This protein was cloned into the pRSET bacterial expression vector for expression in Escherichia coli. In recent years, protein fusion tags have been used to improve recombinant protein expression yields, enable protein purification, and accelerate the characterization of protein structure and function. Immobilized metal-affinity chromatography (IMAC) has been used to purify recombinant proteins that have a polyHistidine (polyHis) tag. Histidine exhibits the strongest interaction with immobilized metal ion matrices due to its electron donor group on the imidazole ring group. We will structurally analyze the purified GCaMP2 using the MS-based protein footprinting method fast photochemical oxidation of proteins (FPOP), which utilizes hydroxyl radicals to oxidatively modify solvent accessible residues of proteins. The hydroxyl radicals are generated by the photolysis of hydrogen peroxide by a pulsed excimer laser. There is a crystal structure available for GCaMP2 which can be used as a model system to test how well FPOP works. For this study, we want to use the FPOP to map the structural differences between calcium-bound and calcium-free form of GCaMP2. We will use EGTA as a chelating agent for the calcium-free form because it has a higher affinity for the calcium. In order to do FPOP on the protein we have to get the protein. So we are expressing in E.coli and purifying it using polyHis tag. We were able to visualize a green distinctive color when the GCaMP2 was purified. We are now able to test how well FPOP works on the protein.

Not Submitted

Funder Acknowledgement(s): Indiana University Historically Black Colleges & Universities STEM Initiative, Jackson State University Louis Stokes Mississippi Alliance for Minority Participation Bridge to the Doctorate

Faculty Advisor: Lisa Jones, joneslis@iupui.edu

Sidebar

Abstract Locators

  • Undergraduate Abstract Locator
  • Graduate Abstract Locator

This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

AAAS

1200 New York Ave, NW
Washington,DC 20005
202-326-6400
Contact Us
About Us

  • LinkedIn
  • Facebook
  • Instagram
  • Twitter
  • YouTube

The World’s Largest General Scientific Society

Useful Links

  • Membership
  • Careers at AAAS
  • Privacy Policy
  • Terms of Use

Focus Areas

  • Science Education
  • Science Diplomacy
  • Public Engagement
  • Careers in STEM

Focus Areas

  • Shaping Science Policy
  • Advocacy for Evidence
  • R&D Budget Analysis
  • Human Rights, Ethics & Law

© 2023 American Association for the Advancement of Science