Title

HIV-1 VIF and human APOBEC3G interaction directly observed through molecular specific labeling using a new dual promotor vector

UMMS Affiliation

Department of Biochemistry and Molecular Biotechnology; Schiffer Lab

Publication Date

2022-04-26

Document Type

Article

Disciplines

Amino Acids, Peptides, and Proteins | Biochemistry | Medicinal Chemistry and Pharmaceutics | Medicinal-Pharmaceutical Chemistry | Molecular Biology | Structural Biology | Virology | Viruses

Abstract

Over the last few decades, protein NMR isotope labeling methods using E. coli based expression have revolutionized the information accessible from biomolecular NMR experiments. Selective labeling of a protein of interest in a multi-protein complex can significantly reduce the number of cross-peaks and allow for study of large protein complexes. However, limitations still remain since some proteins are not stable independently and cannot be separately labeled in either NMR active isotope enriched or unenriched media and reconstituted into a multimeric complex. To overcome this limitation, the LEGO NMR method was previously developed using protein expression plasmids containing T7 or araBAD promoters to separately express proteins in the same E. coli after changing between labeled and unlabeled media. Building on this, we developed a method to label the Human Immunodeficiency Virus type 1 viral infectivity factor (HIV-1 Vif), a monomerically unstable protein, in complex with CBFbeta, it's host binding partner. We designed a dual promoter plasmid containing both T7 and araBAD promoters to independently control the expression of HIV-1 Vif in NMR active isotope enriched media and CBFbeta in unenriched media. Using this method, we assigned the backbone resonance and directly observed the binding of HIV-1 Vif with APOBEC3G, a host restriction factor to HIV-1.

Keywords

APOBEC3G, CBFβ, Double promoter plasmid, HIV-1 Vif, Molecular-specific labeling

DOI of Published Version

10.1016/j.jmr.2022.107230

Source

Myint W, Schiffer CA, Matsuo H. HIV-1 VIF and human APOBEC3G interaction directly observed through molecular specific labeling using a new dual promotor vector. J Magn Reson. 2022 Jun;339:107230. doi: 10.1016/j.jmr.2022.107230. Epub 2022 Apr 26. PMID: 35550909; PMCID: PMC9149140. Link to article on publisher's site

Journal/Book/Conference Title

Journal of magnetic resonance (San Diego, Calif. : 1997)

Related Resources

Link to Article in PubMed

PubMed ID

35550909

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