7QTR

GB1 in mammalian cells, 50 uM

Summary for 7QTR

• Entry DOI: 10.2210/pdb7qtr/pdb
• NMR Information: BMRB: 34700
• Descriptor: Immunoglobulin G-binding protein G (1 entity in total)
• Functional Keywords: gb1, b1 domain of streptococcal protein g, in-cell nmr, immune system
• Biological source: Streptococcus sp. 'group G'
• Total number of polymer chains: 1
• Total formula weight: 6360.00

Authors

Gerez, J.A.,Prymaczok, N.C.,Kadavath, H.,Gosh, D.,Butikofer, M.,Guntert, P.,Riek, R. (deposition date: 2022-01-15, release date: 2022-12-21, Last modification date: 2024-06-19)

Primary citation

Gerez, J.A.,Prymaczok, N.C.,Kadavath, H.,Ghosh, D.,Butikofer, M.,Fleischmann, Y.,Guntert, P.,Riek, R.
Protein structure determination in human cells by in-cell NMR and a reporter system to optimize protein delivery or transexpression.
Commun Biol, 5:1322-1322, 2022

PubMed Abstract: Most experimental methods for structural biology proceed in vitro and therefore the contribution of the intracellular environment on protein structure and dynamics is absent. Studying proteins at atomic resolution in living mammalian cells has been elusive due to the lack of methodologies. In-cell nuclear magnetic resonance spectroscopy (in-cell NMR) is an emerging technique with the power to do so. Here, we improved current methods of in-cell NMR by the development of a reporter system that allows monitoring the delivery of exogenous proteins into mammalian cells, a process that we called here "transexpression". The reporter system was used to develop an efficient protocol for in-cell NMR which enables spectral acquisition with higher quality for both disordered and folded proteins. With this method, the 3D atomic resolution structure of the model protein GB1 in human cells was determined with a backbone root-mean-square deviation (RMSD) of 1.1 Å.

PubMed: 36460747
DOI: 10.1038/s42003-022-04251-6

Experimental method

SOLUTION NMR