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Confocal spectrofluorimetric evidence for the hetero-aggregation of sequence-scrambled forms of two model all-beta sheet proteins.

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title Confocal spectrofluorimetric evidence for the hetero-aggregation of sequence-scrambled forms of two model all-beta sheet proteins.
 
Creator Shukla, Anshuman
Sharma, Swati
Guptasarma, Purnananda
 
Subject QR Microbiology
 
Description Can two unrelated proteins with deliberately compromised folding abilities, marked propensities to aggregate upon increase of protein concentration, and proclivities towards beta sheet formation, be caused to hetero-aggregate? We address this question here using the 'designer' backbone-reversed forms of two model all-beta sheet proteins, E. coli CspA and C. elegans HSP12.6, both earlier created and characterized by our group. These were covalently labeled with fluorescent dyes of well-resolved spectral characteristics [retro-CspA with FITC, and retro-HSP12.6 with TRITC] and then allowed to aggregate within the same reaction vessel. The resultant aggregates are shown by spectrofluorimetry-coupled confocal laser scanning microscopy to constitute uniform mixtures of both proteins, existing within every cylindrical volume element of approximately 200nm diameter, and comparable height, in all sections of the co-aggregated material suggesting that the two proteins do not selectively associate with copies of themselves during aggregation. Thus, it would appear that aggregation can occur without reference to protein molecular identity.
 
Publisher Elsevier Science
 
Date 2007-12-01
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://crdd.osdd.net/open/627/1/guptasarma07.pdf
Shukla, Anshuman and Sharma, Swati and Guptasarma, Purnananda (2007) Confocal spectrofluorimetric evidence for the hetero-aggregation of sequence-scrambled forms of two model all-beta sheet proteins. International journal of biological macromolecules, 41 (5). pp. 650-4. ISSN 0141-8130
 
Relation http://www.sciencedirect.com/science/article/pii/S0141813007001699
http://crdd.osdd.net/open/627/