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Lack of negative charge in the E46Q mutant of photoactive yellow protein prevents partial unfolding of the blue-shifted intermediate.
Derix, Nocky M; Wechselberger, Rainer W; van der Horst, Michael A; Hellingwerf, Klaas J; Boelens, Rolf; Kaptein, Rob; van Nuland, Nico A J.
Afiliação
  • Derix NM; Department of NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Biochemistry ; 42(49): 14501-6, 2003 Dec 16.
Article em En | MEDLINE | ID: mdl-14661962
The long-lived light-induced intermediate (pB) of the E46Q mutant (glutamic acid is replaced by glutamine at position 46) of photoactive yellow protein (PYP) has been investigated by NMR spectroscopy. The ground state of this mutant is very similar to that of wild-type PYP (WT), whereas the pB state, formed upon illumination, appears to be much more structured in E46Q than in WT. The differences are most striking in the N-terminal domain of the protein. In WT, the side-chain carboxylic group of E46 is known to donate its proton to the chromophore upon illumination. The absence of the carboxylic group near the chromophore in the E46Q mutant prohibits the formation of a negative charge at this position upon formation of pB. This prevents the partial unfolding of the mutant, as evidenced from NMR chemical shift comparison and proton/deuterium (H/D) exchange studies.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas de Bactérias / Dobramento de Proteína / Ácido Glutâmico / Fotorreceptores Microbianos / Glutamina / Luz Idioma: En Revista: Biochemistry Ano de publicação: 2003 Tipo de documento: Article País de afiliação: Holanda País de publicação: Estados Unidos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas de Bactérias / Dobramento de Proteína / Ácido Glutâmico / Fotorreceptores Microbianos / Glutamina / Luz Idioma: En Revista: Biochemistry Ano de publicação: 2003 Tipo de documento: Article País de afiliação: Holanda País de publicação: Estados Unidos