Isolation and localization of phosphoprotein pp 135 in the nucleoli of various cell lines
- 1 March 1984
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 139 (2), 417-424
- https://doi.org/10.1111/j.1432-1033.1984.tb08021.x
Abstract
Phosphoprotein pp 135 is one of the dominant proteins endogenously phosphorylated in cellular sonicates during short-time exposure to [.gamma.-32P]ATP. Mouse cells growing exponentially show the highest pp 135 level as determined by endogenous phosphorylation and immunobinding assays. Disruption of cells in the absence of Ca at low Mg concentration renders > 90% pp 135 into the cytosolic fraction. A 5-step purification yields > 95% pure pp 135. The cellular location of pp 135 was determined with a rat anti-(mouse pp 135) serum by immunofluorescence in mouse cell lines and cryostat sections of normal mouse tissue. Fluorescence predominantly of nucleolar structures was observed, confirmed by studies of isolated nuclei and nucleoli. Cross-reacting nucleolar phosphoproteins were identified in cell lines of other species with molecular masses of 128 kDa [kilodalton] (human), 135 kDa (hamster) and 118 kDa (Drosophila). Endogenous phosphorylation of pp 135 investigated with purified mouse nucleoli showed optimal activity at isotonicity, pH 7.3, in the presence of 10 mM Mg ions.This publication has 26 references indexed in Scilit:
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