T proteins of streptococcus pyogenes 4. isolation of t 1 protein by affinity chromatography on immobilized fibrinogen
Schmidt, K.H.; Koehler, W.
Zentralblatt fur Bakteriologie, Mikrobiologie und Hygiene. Series A, Medical Microbiology, Infectious Diseases, Virology, Parasitology 258(4): 449-456
1984
ISSN/ISBN: 0176-6724 Accession: 006577187
T-protein of S. pyrogenes, type 1 (strain SF 130 Griffith) was extracted by enzymatic treatment of the cells with trypsin and partially purified by ion-exchange chromatography on DEAE cellulose and gel chromatography on ultrogel ACA 44. The crude T protein still showing serologically type specific and cross reactions finally was applied to a fibrinogen sepharose column. Components eluted with the neutral buffer (0.5 M phosphate, 0.2 M NaCl, 0.02% NaN3, pH 7.0) reacted serologically in the same manner as the crude T protein. By using 0.1 M citrate,6 M urea pH 3.0 buffer a type specifically reacting protein (T1-TRYP-F) was eluted from the fibrinogen column. T1-TRYP-F showed identical precipitation lines with the recently characterized T1-protein (T1-TRYP-I) purified by immunochromatography on type specific anti-T antibodies. Comparison of the sodium dodecyl sulfate patterns of T1-TRYP-F and T1-TRYP-I revealed a less complex molecular size subunit structure for the fibrinogen binding T1-TRYP-F (2 bands of 60,000 and 70,000) as found for T1-TRYP-I, which showed serologically active peptides .apprx. 30,000-500,000. T protein also may be linked covalently with fibrinogen receptors as was reported for M protein.