A protein isolated from Escherichia coli, identified as GroEL, reacts with anti-beta spectrin antibodies.
We found that a protein of molecular weight close to 65kDa, present in Escherichia coli cells, reacts with anti-beta spectrin antibodies. A method of purification of this protein was designed. The method consists of the following: nonionic detergent extraction, gel filtration chromatography, ion-exchange chromatography using DEAE-Servacell, and two FPLC ion-exchange chromatography runs: the first without urea, the second in its presence. This method allowed us to obtain a highly purified protein. The results of mass spectrometry analysis suggest that the investigated protein is GroEL (Hsp60 Class). Using computer programs, by sequence analysis of both proteins we tried to explain why GroEL isolated from E. coli reacts with anti-beta spectrin antibodies. Both proteins may share a single epitope for the antibodies on their surfaces. Additionally, such an assumption is supported by the results of experiments in which antibodies interacting with GroEL were obtained from anti-beta spectrin serum and were shown to react with both GroEL and beta spectrins.
Czogalla
Aleksander
A
Institute of Biochemistry and Molecular Biology, University of Wrocław, ul. Przybyszewskiego 63/77, 51-148 Wrocław, Poland.
Kwolek
Paulina
P
Hryniewicz-Jankowska
Anita
A
Nietubyć
Małgorzata
M
Leluk
Jacek
J
Sikorski
Aleksander F
AF
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism; D000915:Antibody Affinity; D000918:Antibody Specificity; D001483:Base Sequence; D018834:Chaperonin 60 / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002845:Chromatography / Q000379:methods; D000939:Epitopes / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D008970:Molecular Weight; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment / Q000379:methods; D020539:Sequence Analysis, Protein / Q000379:methods; D013045:Species Specificity; D013049:Spectrin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
D000911:Antibodies, Monoclonal; D018834:Chaperonin 60; D000939:Epitopes; D021122:Protein Subunits; D013049:Spectrin