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Doctoral Thesis
DOI
https://doi.org/10.11606/T.46.2007.tde-15082007-074551
Document
Author
Full name
Vitor Renaux Hering
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2007
Supervisor
Committee
Alário, Adelaide Faljoni (President)
Feitosa, Eloi da Silva
Mammana, Victor Pellegrini
Politi, Mário José
Schumacher, Robert
Title in Portuguese
Interação entre proteínas fluorescentes e nanocristais de CdSe/ZnS
Keywords in Portuguese
CdSe/ZnS
FRET
GFP
HcRed1
Nanocristais
ProteÌnas fluorescentes
Abstract in Portuguese
Foram utilizadas proteínas da famÌlia das GFPs e nanocristais fluorescentes de CdSe/ZnS para caracterização da interação e verificação de transferência de energia por ressonância (FRET) entre estes compostos. Formou-se dois pares doador-receptor onde ora uma proteína figurava como doadora, ora um nanocristal ocupava este papel. Verificou-se que, em ambos os casos, o doador sofre supressão da fluorescência após a formação de complexo com o receptor, complexo este motivado por interação eletrostática e dependente de pH. Foi possível comprovar, através da observação de emissão sensitizada e redução da anisotropia, que entre o par formado por nanocristal com emissão no verde e proteína HcRed1 como receptora, de fato ocorre FRET. As distâncias aparentes entre doador e receptor foram determinadas a partir da eficiência da supressão da fluorescência do doador e da distância de Förster. As distâncias assim obtidas são compatíveis com as dimensões das proteínas e dos nanocristais
Title in English
Interaction between fluorescent proteins and CdSe/ZnS nanocrystals
Keywords in English
CdSe/ZnS
Fluorescent proteins
FRET
GFP
HcRed1
Nanocrystals
Abstract in English
Proteins belonging to the GFP family were used to characterize their interaction with fluorescent CdSe/ZnS nanocrystals and to verify the occurrence of resonance energy transfer (FRET) among these elements. Two donor-acceptor pairs were established, one having a protein as donor and the other having a nanocrystal as donor. In both cases the donor suffers quenching of its fluorescence after the formation of a complex with the acceptor. The complex formation is dependent on pH and is due to electrostatic interaction. It was possible to prove the occurrence of FRET between CdSe/ZnS nanocrystals emitting green fluorescence as donors and the protein HcRed1 as acceptor, through the detection of sensitized emission and anisotropy reduction. Apparent donor-acceptor distances were determined from efficiency measurements and Förster distances. The obtained distances agreed with the protein and nanocrystal dimensions
 
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ErrataVitorRenaux.pdf (11.03 Kbytes)
TESEVITORRENAUX.pdf (764.52 Kbytes)
Publishing Date
2007-09-17
 
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