Two-component atomic force microscopy recognition imaging of complex samples

Biological complexes are typically multisubunit in nature and the processes in which they participate often involve protein compositional changes in themselves and/or their target substrates. Being able to identify more than one type of protein in complex samples and to track compositional changes d...

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Bibliographic Details
Published in:Analytical biochemistry 2007-02, Vol.361 (2), p.273-279
Main Authors: Wang, H., Bash, R., Lohr, D.
Format: Article
Language:English
Subjects:
Actins - analysis
AFM
Chromatin
Chromatin - chemistry
Chromosomal Proteins, Non-Histone - chemistry
DNA - chemistry
DNA Helicases - analysis
Histones - analysis
hSwi–Snf
Humans
Microscopy, Atomic Force - methods
Multiprotein Complexes - chemistry
Nuclear Proteins - analysis
Nucleosomal arrays
Proteins - analysis
Recognition imaging
SPM
Transcription Factors - analysis
Transcription Factors - chemistry
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Summary:Biological complexes are typically multisubunit in nature and the processes in which they participate often involve protein compositional changes in themselves and/or their target substrates. Being able to identify more than one type of protein in complex samples and to track compositional changes during processes would thus be very useful. Toward this goal, we describe here a single-molecule technique that can simultaneously identify two types of proteins in compositionally complex samples. It is an adaptation of the recently developed atomic force microscopy (AFM) recognition imaging technique but involves the tethering of two different types of antibodies to the AFM tip and sequential blocking with appropriate antigenic peptides to distinguish the recognition from each antibody. The approach is shown to be capable of simultaneously identifying in a single AFM image two specific components, BRG1 and β-actin, of the human Swi–Snf ATP-dependent nucleosome remodeling complex and two types of histones, H2A and H3, in chromatin samples.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2006.11.039