Gelation and fodrin purification from rat brain extracts.
Extracts from rat brain tissue have been shown to give rise to a gel which exhibits the following features. It is mainly enriched in actin and in a high-molecular-weight protein with polypeptide chains of 235 and 240 kDa, which we identified as fodrin. Tubulin is also a major component of the gel but it appears to be trapped non-specifically during the gelation process. Gelation is pH-, ionic strength- and Ca2+-concentration-dependent, and is optimal under the conditions which promote the interaction between polymerized actin and fodrin. In a similar way to that described for the purification of rat brain actin (Levilliers, N., Péron-Renner, M., Coffe, G. and Pudles, J. (1984) Biochimie 66, 531-537), we used the gelation system as a selective means of recovering fodrin from the mixture of a low-ionic-strength extract from whole rat brain and a high-ionic-strength extract of the particulate fraction. From this gel, fodrin was purified with a good yield by a simple procedure involving gel dissociation in 0.5 M KCl and depolymerization in 0.7 M KI, Bio-Gel A-15m chromatography, followed by ammonium sulfate precipitation.
N
N
Levilliers
M
M
Péron-Renner
G
G
Coffe
J
J
Pudles
D000199:Actins / Q000302:isolation & purification; D000818:Animals; D001923:Brain Chemistry; D002118:Calcium / Q000502:physiology; D002352:Carrier Proteins / Q000302:isolation & purification*; D005260:Female; D005782:Gels / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008840:Microfilament Proteins / Q000302:isolation & purification*; D009994:Osmolar Concentration; D051381:Rats; D011919:Rats, Inbred Strains; D014020:Tissue Extracts; D014404:Tubulin / Q000302:isolation & purification; D013049:Spectrin
D000199:Actins; D002118:Calcium; D002352:Carrier Proteins; D005782:Gels; D008840:Microfilament Proteins; D014020:Tissue Extracts; D014404:Tubulin; D013049:Spectrin