DNA, oligosaccharides, and mononucleotides stimulate oligomerization of human lactoferrin.
Using small-angle X-ray scattering (SAXS), light scattering (LS), and soft laser ablation we have shown that lactoferrin (LF) in solution at neutral pH is oligomerized in the absence of salt or at physiological salt concentrations. The level of oligomerization depends on the concentration of LF, KCl or NaCl, and on the duration of the protein storage in solution. At the concentrations comparable with those in human milk (1-6 mg/ml), the average radius of gyration (R(g)) values of LF can attain 400-480 A, while fresh solution of previously lyophylized LF demonstrate a lower average R(g) (50-100 A), and R(g) value characterizing the LF monomer formed at 1 M NaCl is 26.7 A. The addition of oligonucleotides, oligosaccharides, or mononucleotides to LF in the presence or in the absence of KCl with different level of initial oligomerization accelerates the oligomerization rate and increases the R(g) values up to approximately 600-700 A, which correspond to associates containing ten or more protein molecules. During gel filtration on Sepharose 4B, high-degree LF oligomers dissociate nearly completely forming different degraded complexes, but in some cases it is possible to reveal small amount of a decamer. A possible role for oligomerization of LF, a highly polyfunctional protein, for its different biological activities is discussed.
Nevinskii
Alexander G
AG
Institute of Chemical Biology and Fundamental Medicine, Siberian Division of the Russian Academy of Sciences, Novosibirsk, Russia.
Soboleva
Svetlana E
SE
Tuzikov
Fedor V
FV
Buneva
Valentina N
VN
Nevinsky
Georgy A
GA
D002850:Chromatography, Gel; D004247:DNA / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D007781:Lactoferrin / Q000737:chemistry*; D007834:Lasers; D008024:Ligands; D009711:Nucleotides / Q000494:pharmacology*; D009844:Oligosaccharides / Q000494:pharmacology*; D020836:Protein Structure, Quaternary / Q000187:drug effects; D053838:Scattering, Small Angle; D012996:Solutions; D013997:Time Factors; D017186:Titrimetry; D014961:X-Ray Diffraction
D008024:Ligands; D009711:Nucleotides; D009844:Oligosaccharides; D012996:Solutions; D004247:DNA; D007781:Lactoferrin