On the effect of sodium dodecyl sulfate on the structure of beta-galactosidase from Escherichia coli. A fluorescence study.
An understanding of the structure-function relationship of proteins under different chemical-physical conditions is of fundamental importance for an understanding of their structure and function in cells. In this paper we report the effects of sodium dodecyl sulfate and temperature on the structure of beta-galactosidase from Escherichia coli, as monitored by fluorescence spectroscopy. The structure of the protein was studied in the temperature range of 10-60 degrees C in the absence and presence of sodium dodecyl sulfate by frequency-domain measurement of the intrinsic fluorescence intensity and anisotropy decays. The time-resolved fluorescence data in the absence of SDS indicated that at 10 degrees C the tryptophanyl emission decays were well described by a three exponential decays model, and that the temperature increase resulted in shortening of the long-lived component with little change in the short- and middle-lived components. The addition of SDS to the protein solution also affected the long-lived component. The effects of the detergent and temperature on the enzyme structure were also investigated by means of quenching experiments and anisotropy decays. The obtained results showed that the presence of SDS confers more flexibility to the protein structure, and suggest a strict relation between enzyme activity and protein flexibility.
D'Auria
S
S
Center for Fluorescence Spectroscopy, University of Maryland at Baltimore, West Lombard Street, Baltimore, MD 21201 USA. sabato@cfs.umbi.umd.edu
Di Cesare
N
N
Gryczynski
I
I
Rossi
M
M
Lakowicz
J R
JR
D020106:Acrylamide / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology*; D005454:Fluorescence Polarization; D008958:Models, Molecular; D017434:Protein Structure, Tertiary / Q000187:drug effects; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D013696:Temperature; D014364:Tryptophan / Q000737:chemistry; D001616:beta-Galactosidase / Q000737:chemistry*
D020106:Acrylamide; D012967:Sodium Dodecyl Sulfate; D014364:Tryptophan; D001616:beta-Galactosidase