Binding of glutamine to glutamine-binding protein from Escherichia coli induces changes in protein structure and increases protein stability.
Glutamine-binding protein (GlnBP) from Escherichia coli is a monomeric protein localized in the periplasmic space of the bacterium. It is responsible for the first step in the active transport of L-glutamine across the cytoplasmic membrane. The protein consists of two similar globular domains linked by two peptide hinges, and X-ray crystallographic data indicate that the two domains undergo large movements upon ligand binding. Fourier transform infrared spectroscopy (FTIR) was used to analyze the structure and thermal stability of the protein in detail. The data indicate that glutamine binding induces small changes in the secondary structure of the protein and that it renders the structure more thermostable and less flexible. Detailed analyses of IR spectra show a lower thermal sensitivity of alpha-helices than beta-sheets in the protein both in the absence and in the presence of glutamine. Generalized two-dimensional (2D) analyses of IR spectra reveal the same sequence of unfolding events in the protein in the absence and in the presence of glutamine, indicating that the amino acid does not affect the unfolding pathway of the protein. The data give new insight into the structural characteristics of GlnBP that are useful for both basic knowledge and biotechnological applications.
D'Auria
Sabato
S
Institute of Protein Biochemistry, CNR, Via Pietro Castellino, 111 Naples, Italy. s.dauria@ibp.cnr.it <s.dauria@ibp.cnr.it>
Scirè
Andrea
A
Varriale
Antonio
A
Scognamiglio
Viviana
V
Staiano
Maria
M
Ausili
Alessio
A
Marabotti
Anna
A
Rossi
Mosè
M
Tanfani
Fabio
F
D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D005973:Glutamine / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D017433:Protein Structure, Secondary / Q000502:physiology; D013816:Thermodynamics
D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C037853:glutamine transport proteins; D005973:Glutamine