AFM visualization of clathrin triskelia under fluid and in air

Kotova, Svetlana; Prasad, Kondury; Smith, Paul D.; Lafer, Eileen M.; Nossal, Ralph; Jin, Albert J.

doi:10.1016/j.febslet.2009.11.039

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Volume 584, Issue 1 , Pages 44-48, 4 January 2010

AFM visualization of clathrin triskelia under fluid and in air

Edited by Michael R. Bubb

Svetlana Kotova
- Laboratory of Bioengineering and Physical Science, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, DHHS, Bethesda, MD 20892, United States
Kondury Prasad
- Department of Biochemistry, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, United States
Paul D. Smith
- Laboratory of Bioengineering and Physical Science, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, DHHS, Bethesda, MD 20892, United States
Eileen M. Lafer
- Department of Biochemistry, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, United States
Ralph Nossal
- Laboratory of Integrative and Medical Biophysics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, DHHS, Bethesda, MD 20892, United States
- Corresponding authors. Fax: +1 301 496 6608 (A.J. Jin).
Albert J. Jin
- Laboratory of Bioengineering and Physical Science, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, DHHS, Bethesda, MD 20892, United States
- Corresponding authors. Fax: +1 301 496 6608 (A.J. Jin).

Received 3 September 2009; received in revised form 9 November 2009; accepted 10 November 2009. published online 16 November 2009.

Abstract

Atomic force microscopy (AFM) is used to characterize the structure and interactions of clathrin triskelia. Time sequence images of individual, wet triskelia resting on mica surfaces clearly demonstrate conformational fluctuations of the triskelia. AFM of dried samples yields images having nanometric resolution comparable to that obtainable by electron microscopy of shadowed samples. Increased numbers of triskelion dimers and assembly intermediates, as well as structures having dimensions similar to those of clathrin cages, are observed when the triskelia were immersed in a low salt, low pH buffer. These entities have been quantified by AFM protein volume computation.