Preparation and characterization of myosin from cultured cells of Escherichia coli (02:K1 30156 strain).
In this study the myosin preparations isolated from E. Coli cell cultures were analysed. The isolation of myosin from fresh cultures resulted in a substantially higher (approximately 10-fold) yield than from stored cells. The coli myosin, despite the two DEAE-cellulose treatments, contained more RNA and P-lipid than the myosin prepared from skeletal muscle. The RNA content can be removed gradually by acetone denaturation and lipid removal followed by subsequent washings. The fresh preparations contained protein-bound alkali-stable P; part of this was released by Cu ions. The coli myosin can be phosphorylated. The phosphate uptake depended on the concentrations of ATP and Mg2+ and on the time of incubation. The alkaline hydrolysate of the phosphorylated, washed lipid-free preparation was resolved into 8 P-containing peaks on ion-exchange chromatography. Of fractions P-Arg, Pi, P-Lys and 2 P-His conformer were identified by means of synthetic compounds, elution pattern and specific reactions. The remaining compounds could not be identified. The most abundant component was P-Arg suggesting that this compound might play an important role in the cytokinetic movements of E. coli.
Fazekas
S
S
Solymosi
M
M
Ováry
I
I
Székessy-Hermann
V
V
Antoni
F
F
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000596:Amino Acids / Q000032:analysis; D002478:Cells, Cultured; D005591:Chemical Fractionation; D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel; D004926:Escherichia coli / Q000032:analysis*; D006868:Hydrolysis; D009218:Myosins / Q000032:analysis / Q000302:isolation & purification*; D010710:Phosphates / Q000032:analysis; D010766:Phosphorylation; D012313:RNA / Q000032:analysis
D000596:Amino Acids; D010710:Phosphates; D012313:RNA; D000255:Adenosine Triphosphate; D009218:Myosins