Effect of pH on the structure and aggregation of human glycodelin A. A comparison with beta-lactoglobulin A.
The effect of pH on the structure of glycodelin A (GdA) and of beta-lactoglobulin A (beta-LgA) has been investigated by means of circular dichroism, steady state fluorescence, synchrotron radiation small angle X-ray scattering (SR-SAXS) and gel permeation chromatography. The comparison between GdA and beta-LgA shows that, at pH 7.0, both proteins are dimers with an extended content of beta-sheet conformation, but pH 2.0 and 9.0 yield a different secondary, tertiary and quaternary structural organisation. Whilst beta-LgA is a monomer, that conserves beta-sheet conformation at pH 2.0 and 9.0, GdA has a stable dimeric structure at alkaline pH, but at pH 2.0 increases its alpha-helix content and it aggregates soon. SR beam has been used to perform SAXS comparative measurements of the two proteins. SR-SAXS data provide the radius of gyration and the radii of the cross-section and of the thickness. GdA aggregation at acid pH has been characterised by calculating the distance distribution function (P(r)). Isoelectric focusing and chromatofocusing data show a different charge distribution on the surfaces of the two proteins, supporting the hypothesis that the presence of oligosaccharides deeply influences the conformational state and the aggregation process of GdA at different pH values. In particular, the presence of sialic acid residues, within the oligosaccharide moiety of the GdA, might be responsible for the differences observed between the two proteins.
Barteri
M
M
Dipartimento di Chimica, Università degli Studi La Sapienza, Rome, Italy. barteri@caspur.it
Gaudiano
M C
MC
Rotella
S
S
Benagiano
G
G
Pala
A
A
D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D000071816:Glycodelin; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007782:Lactoglobulins / Q000737:chemistry*; D011257:Pregnancy Proteins / Q000737:chemistry*; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D000071816:Glycodelin; D006023:Glycoproteins; D007782:Lactoglobulins; C045246:PAEP protein, human; D011257:Pregnancy Proteins