Methods for Crystallization and Structural Determination of M-T7 Protein from Myxoma Virus

Christopher Gisriel, Petra Fromme, Jose M. Martin-Garcia

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The myxoma virus has become of interest in human medicine in the last two decades as it has the ability to infect many types of human cancer cells and is being used as a platform to develop viro-therapeutic agents that suppress aggressive and damaging immune responses and inflammation. Furthermore, the myxoma virus encodes proteins that have strong immunosuppressive effects, and several of the myxoma virus-encoded immunomodulators are being developed to treat systemic inflammatory syndromes such as cardiovascular disease and transplant rejection. Myxoma virus encodes the M-T7 protein, the most abundantly secreted protein expressed in myxoma virus-infected cells, originally identified as a rabbit species-specific interferon-gamma (IFN-γ) receptor homolog and as a chemokine-modulating protein binding a wide range of mammalian chemokines. M-T7 is a critical virulence factor for viral pathogenesis that increases virus lethality when expressed. Although M-T7 has been extensively studied using biochemical and biophysical techniques and its interactome map is well known, its three-dimensional (3D) structure remains elusive. Obtaining the 3D structure of M-T7 would be greatly beneficial and is a crucial step toward advancing M-T7 research through understanding the molecular function and activity of M-T7 as a novel therapeutic reagent and to rationally develop this protein as a drug. This chapter provides an overview of the structural determination techniques, especially X-ray crystallography, that can be applied toward the goal of achieving the first high-resolution structure of M-T7. In addition, details of up-and-coming methods are discussed, including X-ray diffraction at X-ray free electron lasers (XFELs), nuclear magnetic resonance (NMR), cryo-electron microscopy (cryo-EM), Micro-electron diffraction (Micro-ED), and small-angle X-ray scattering (SAXS), and their potential applications to M-T7 structural biology.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages125-162
Number of pages38
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2225
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cryo-electron microscopy (cryo-EM)
  • M-T7 protein
  • Micro-electron diffraction (Micro-ED)
  • Myxoma virus
  • Small-angle X-ray scattering (SAXS)
  • Structure determination
  • X-ray crystallography
  • X-ray free electron lasers (XFELs)

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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