Species-dependent binding of serum albumins to the streptococcal receptor protein G.
The interaction of the serum albumin binding domain from streptococcal protein G to serum albumins isolated from different species was investigated. The highest affinity to protein G was found for serum albumins from rat, man and mouse. A medium binding was found for serum albumin from rabbit, cow, hen and horse, while little or no binding was found for ovalbumin and serum albumin from sheep. The interaction between human serum albumin and protein G showed rapid binding kinetics at the temperatures 7, 22 and 37 degrees C. Furthermore, the ability of different serum albumins to function as affinity ligands when covalently coupled to a solid support was tested. The results show that protein G derivatives could be eluted at different pH depending on the origin of the serum albumin. It was also possible to elute the streptococcal receptor efficiently from the mouse serum albumin matrix with human serum albumin. Based on these results, a gene fusion system for recovery of sensitive proteins by affinity purification is described, where high yields are obtained under mild elution conditions.
Nygren
P A
PA
Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden.
Ljungquist
C
C
Trømborg
H
H
Nustad
K
K
Uhlén
M
M
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D001667:Binding, Competitive; D002417:Cattle; D003001:Cloning, Molecular; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D051379:Mice; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011817:Rabbits; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism*; D013045:Species Specificity; D013329:Structure-Activity Relationship
D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D009838:Oligodeoxyribonucleotides; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D012709:Serum Albumin