Type II CaS /calmodulin-dependent kinase phosphorylates tau protein in the region of the mouse repeat
Tau is a phosphoprotein associated with the subset of microtubules present in the axonal domain of neurons and is a component of the Alzheimer neurofibrillary tangle. Tau protein was purified from bovine brain by the taxol method followed by gel filtration of the heat-stable microtubule fraction CaS /calmodulin-dependent kinase was purified from rat brain as previously described. Incubation of tau in the kinase-containing reaction mixture resulted in intense incorporation of TSP into the protein. Labeled tau protein was trypsinized and separated into discrete fragments by reverse-phase HPLC. The chromatogram contained two radioactive peaks, sequenced on a gas phase sequenator. The first peak was eluted into two tubes, with more radioactivity in the second tube. The sequence in the second tube contained 16 residues which corresponded to an identical sequence in mouse tau that is part of a longer stretch which repeats three times with greater than 50% homology. The highest CPM's were located on a PTH-conjugated threonine. The first tube contained the identical amino acid sequence and a lower level of radioactivity throughout unassociated with any single amino acid. About 1/3 of the total peptide was phosphorylated. A second radioactive peak contained an unidentifiable PTH-amino acid present at a level below the yield obtained for tau.
- Research Organization:
- Harvard Medical School, Boston, MA
- OSTI ID:
- 6156363
- Report Number(s):
- CONF-870644-; TRN: 87-028703
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
- Country of Publication:
- United States
- Language:
- English
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PHOSPHOTRANSFERASES
FRACTIONATION
PROTEINS
LABELLING
PHOSPHORYLATION
AMINO ACID SEQUENCE
CALCIUM COMPOUNDS
CATIONS
LIQUID COLUMN CHROMATOGRAPHY
MICE
NERVES
PHOSPHORUS 32
TRACER TECHNIQUES
ALKALINE EARTH METAL COMPOUNDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHARGED PARTICLES
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ENZYMES
IONS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MOLECULAR STRUCTURE
NERVOUS SYSTEM
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
RADIOISOTOPES
RODENTS
SEPARATION PROCESSES
TRANSFERASES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques