Fabs enable single particle cryoEM studies of small proteins

Shenping Wu, Agustin Avila-Sakar, Jungmin Kim, David S. Booth, Charles H. Greenberg, Andrea Rossi, Maofu Liao, Xueming Li, Akram Alian, Sarah L. Griner, Narinobu Juge, Yadong Yu, Claudia M. Mergel, Javier Chaparro-Riggers, Pavel Strop, Robert Tampé, Robert H. Edwards, Robert M. Stroud, Charles S. Craik, Yifan Cheng

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

In spite of its recent achievements, the technique of single particle electron cryomicroscopy (cryoEM) has not been widely used to study proteins smaller than 100 kDa, although it is a highly desirable application of this technique. One fundamental limitation is that images of small proteins embedded in vitreous ice do not contain adequate features for accurate image alignment. We describe a general strategy to overcome this limitation by selecting a fragment antigen binding (Fab) to form a stable and rigid complex with a target protein, thus providing a defined feature for accurate image alignment. Using this approach, we determined a three-dimensional structure of an ∼65 kDa protein by single particle cryoEM. Because Fabs can be readily generated against a wide range of proteins by phage display, this approach is generally applicable to study many small proteins by single particle cryoEM.

Original languageEnglish
Pages (from-to)582-592
Number of pages11
JournalStructure
Volume20
Issue number4
DOIs
Publication statusPublished - Apr 4 2012
Externally publishedYes

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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    Wu, S., Avila-Sakar, A., Kim, J., Booth, D. S., Greenberg, C. H., Rossi, A., Liao, M., Li, X., Alian, A., Griner, S. L., Juge, N., Yu, Y., Mergel, C. M., Chaparro-Riggers, J., Strop, P., Tampé, R., Edwards, R. H., Stroud, R. M., Craik, C. S., & Cheng, Y. (2012). Fabs enable single particle cryoEM studies of small proteins. Structure, 20(4), 582-592. https://doi.org/10.1016/j.str.2012.02.017