Denaturation and partial renaturation of a tightly tetramerized DsRed protein under mildly acidic conditions.

Abstract:

:The red fluorescent protein, DsRed, recently cloned from coral Discosoma sp. has one of the longest fluorescence waves and one of the most complex absorbance spectra among the family of fluorescent proteins. In this work we found that with time DsRed fluorescence decreases under mildly acidic conditions (pH 4.0-4.8) in a pH-dependent manner, and this fluorescence inactivation could be partially recovered by subsequent re-alkalization. The DsRed absorbance and circular dichroism spectra under these conditions revealed that the fluorescence changes were caused by denaturation followed by partial renaturation of the protein. Further, analytical ultracentrifugation determined that native DsRed formed a tight tetramer under various native conditions. Quantitative analysis of the data showed that several distinct states of protein exist during the fluorescence inactivation and recovery, and the inactivation of fluorescence can be caused by protonation of a single ionogenic group in each monomer of DsRed tetramer.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Vrzheshch PV,Akovbian NA,Varfolomeyev SD,Verkhusha VV

doi

10.1016/s0014-5793(00)02344-9

keywords:

subject

Has Abstract

pub_date

2000-12-29 00:00:00

pages

203-8

issue

2

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(00)02344-9

journal_volume

487

pub_type

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