The binding of a putative and a known chaperone protein revealed by immunogold labelling transmission electron microscopy: A suggested use of chaperones as probes for the distribution of their target proteins

Thorpe, Julian R, Rulten, Stuart L and Kay, John E (1999) The binding of a putative and a known chaperone protein revealed by immunogold labelling transmission electron microscopy: A suggested use of chaperones as probes for the distribution of their target proteins. Journal of Histochemistry and Cytochemistry, 47. pp. 1633-1640.

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Abstract

Parvulins are a distinct family within the peptidyl-prolyl cis-trans isomerase group of proteins that catalyse the cis-trans isomerization of proline-containing peptides. The intracellular distribution of a novel human parvulin homologue (hEPVH) has been investigated in a human kidney cell line (HEK 293) by immunogold labeling transmission electron microscopy (TEM). This showed hEPVH to be distributed throughout HEK 293 cells but in highest concentration within mitochondria. Unexpectedly, preabsorption of anti-hEPVH antiserum with recombinant hEPVH exaggerated the observed immunolabel density in a pattern that mirrored that of the endogenous hEPVH. The hEPVH protein itself was then used to label sections, and the specificity of its binding was confirmed with the use of polyclonal and monoclonal antibodies in conjunction with homologous and irrelevant protein controls. The pattern of hEPVH binding also mirrored that of endogenous hEPVH. A known chaperone protein, Pin1, was also found to bind to cells in a pattern mirroring that of the endogenous protein. This lends considerable weight to our hypothesis that hEPVH is binding to its target protein(s) within the cell, reflecting its postulated chaperone function. Finally, we suggest that chaperone proteins in general might be used as TEM probes for their target (or substrate) proteins. (J Histochem Cytochem 47:1633-1640, 1999)

Item Type: Article
Schools and Departments: School of Life Sciences > Biochemistry
Depositing User: Julian Thorpe
Date Deposited: 06 Feb 2012 19:56
Last Modified: 22 Mar 2012 09:30
URI: http://sro.sussex.ac.uk/id/eprint/23120
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