Partial Characterization of Endogenous Phosphorylation Conditions for Hen Brain Cytosolic and Membrane Proteins

Details

• Personal Author:
  Abou-Donia MB ; Lapadula DM ; Patton SE
• Description:
  The optimal conditions for endogenous protein phosphorylation with 5uM [gamma-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 75-125ug/200 u1 in brain cytosol and was maximal at 75 ug/200 ul 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 45 degrees C. 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 µM 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. [Description provided by NIOSH]
• Subjects:
  Animals Environmental Monitoring Laboratories Nervous System Nervous System Diseases Occupational Health Safety
• Keywords:
  Animal-studies Author Keywords: Brain Cytosol Brain-damage Brain-function Brain Membranes Central-nervous-system Exposure-assessment Exposure-levels Laboratory-animals Neurotoxic-effects Neurotoxicity Protein-biochemistry Protein Phosphorylation Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis

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• ISSN:
  0006-8993
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  North Carolina ; OSHA Region 4
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  1-14
• Volume:
  328
• Issue:
  1
• NIOSHTIC Number:
  nn:20037433
• Citation:
  Brain Res 1985 Feb; 328(1):1-14
• Federal Fiscal Year:
  1985
• NORA Priority Area:
  Neurotoxic Effects
• Performing Organization:
  Duke University, Durham, North Carolina
• Peer Reviewed:
  True
• Start Date:
  19840601
• Source Full Name:
  Brain Research
• End Date:
  19881031
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:f5880c97c5c90d15bcf5f84ed7b88cfff6fc5e8ad91d918056f2cfdc1e1ed8795ebcb0f989e09025beb923af94fbff067ae2b010315f1a4c9b0908f57f9159d2
• Download URL:
  https://stacks.cdc.gov/view/cdc/188530/cdc_188530_DS1.pdf
• File Type:
  [PDF - 2.54 MB ]