Skip directly to search Skip directly to A to Z list Skip directly to page options Skip directly to site content

NIOSHTIC-2 Publications Search

Search Results

Partial characterization of endogenous phosphorylation conditions for hen brain cytosolic and membrane proteins.

Authors
Patton-SE; Lapadula-DM; Abou-Donia-MB
Source
Brain Res 1985 Feb; 328(1):1-14
NIOSHTIC No.
20037433
Abstract
The optimal conditions for endogenous protein phosphorylation with 5M [Y-32P]ATP, 10 mM MgCl2 in preparations containing synaptosomal cytosol or membranes (shocked crude mitochondrial fraction P2) from adult hen brains were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, autoradiography and microdensitometry. Phosphate incorporation increased linearly with protein concentration from 75-125g/200 1 in brain cytosol and was maximal at 75 g/200 l in brain membranes. Optimal incubation times were 60-90s for brain cytosol and 10-15 s for brain membranes. With the exception of the 20 kilodalton myelin basic protein in the membrane fraction, pH 6.5 is generally optimal. While temperature optima varied considerably with different bands, most of them were found between 35 and 45C. When identical preparations from hen and rat brain were co-electrophoresed, one of the most striking differences was that the enhancement of phosphorylation of a 55 kilodalton doublet, which may be tubulin, by addition of 50 M Ca2+ was at least 3 times greater in rat than in hen brain cytosol. Another species difference was apparent in the membrane fractions in which the 20 kilodalton hen brain persumptive myelin basic protein (MBP) was phosphorylated to approximately the same extent as that of the 16 and 20 kilodalton rat brain MBPs combined.
Keywords
Neurotoxic-effects; Protein-biochemistry; Neurotoxicity; Exposure-assessment; Exposure-levels; Central-nervous-system; Animal-studies; Laboratory-animals; Brain-damage; Brain-function; Author Keywords: brain cytosol; brain membranes; protein phosphorylation; sodium dodecyl sulfate-polyacrylamide gel electrophoresis
CODEN
BRREAP
Publication Date
19850225
Document Type
Journal Article
Funding Amount
665055
Funding Type
Grant
Fiscal Year
1985
NTIS Accession No.
NTIS Price
Identifying No.
Grant-Number-R01-OH-02003; Grant-Number-R01-OH-00823
Issue of Publication
1
ISSN
0006-8993
Priority Area
Neurotoxic-effects
Source Name
Brain Research
State
NC
Performing Organization
Duke University, Durham, North Carolina
TOP