Evidence for a high molecular weight cytosolic factor that binds brain and liver metallothionein.
When brain extracts were fractionated in a Sephadex G-75 chromatography and MT levels were assayed by RIA or ELISA using polyclonal antibodies specific for the MT-I and MT-II isoforms, it was found that MT mostly eluted in the high molecular weight (HMW) peak even in reducing or anaerobic conditions. This was also the case for the liver extracts of control rats; in stressed animals MT immunoreactivity in the HMW peak (> 80 Kd) was increased compared with undisturbed animals, but the major amount of the newly induced MT eluted, as expected from the current literature, in the low molecular weight (LMW) peak, around 10 Kd. The addition of purified MT to brain extracts precluded its binding to a DEAE-Sephadex column. Furthermore, immunoblot results of native PAGE showed that MT changed its electrophoretic mobility in the presence of HMW proteins from brain cytosol. Altogether, these results suggest that a cytosolic factor binds MT in a saturable manner, which may have strong physiological implications.
Department de Biologia Cellular i de Fisiologia, Facultat de Ciències, Universitat Autònoma de Barcelona, Bellaterra, Spain.
T
T
Gasull
J
J
Hidalgo
D000693:Anaerobiosis; D000818:Animals; D000918:Antibody Specificity; D001921:Brain / Q000378:metabolism*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D015151:Immunoblotting; D008099:Liver / Q000378:metabolism*; D008668:Metallothionein / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats; D015032:Zinc / Q000494:pharmacology
D011506:Proteins; D008668:Metallothionein; D015032:Zinc