Development of a novel photoreactive calmodulin derivative: Cross-linking of purified adenylate cyclase from bovine brain
- Univ. of Michigan, Ann Arbor (USA)
A novel photoreactive calmodulin (CaM) derivative was developed and used to label the purified CaM-sensitive adenylate cyclase from bovine cortex. {sup 125}I-CaM was conjugated with the heterobifunctional cross-linking agent p-nitrophenyl 3-diazopyruvate (DAPpNP). Spectral data indicated that diazopyruvoyl (DAP) groups were incorporated into the CaM molecule. Iodo-CaM-DAPs behaved like native CaM with respect to (1) Ca{sup 2+}-dependent enhanced mobility on sodium dodecyl sulfate-polyacrylamide gels and (2) Ca{sup 2+}-dependent stimulation of adenylate cyclase activity. {sup 125}I-CaM-DAP photochemically cross-linked to CaM-binding proteins in a manner that was both Ca{sup 2+} dependent and CaM specific. Photolysis of forskolin-agarose-purified adenylate cyclase from bovine cortex with {sup 125}I-CaM-DAP produced a single cross-linked product which migrates on sodium dodecyl sulfate-polyacrylamide gels with an apparent molecular weight of approximately 140,000.
- OSTI ID:
- 7018934
- Journal Information:
- Biochemistry; (USA), Vol. 28:14; ISSN 0006-2960
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
BRAIN
CROSS-LINKING
CALMODULIN
DERIVATIZATION
CYCLASES
ATP
AUTORADIOGRAPHY
CALCIUM CHLORIDES
ENZYME ACTIVITY
IODINE 125
PHOSPHORUS 32
PROTEINS
SODIUM IODIDES
TRITIUM COMPOUNDS
ALKALI METAL COMPOUNDS
ALKALINE EARTH METAL COMPOUNDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CALCIUM COMPOUNDS
CALCIUM HALIDES
CENTRAL NERVOUS SYSTEM
CHEMICAL REACTIONS
CHLORIDES
CHLORINE COMPOUNDS
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
HALIDES
HALOGEN COMPOUNDS
HYDROGEN COMPOUNDS
INORGANIC PHOSPHORS
INTERMEDIATE MASS NUCLEI
IODIDES
IODINE COMPOUNDS
IODINE ISOTOPES
ISOTOPES
LIGHT NUCLEI
LYASES
NERVOUS SYSTEM
NUCLEI
NUCLEOTIDES
ODD-EVEN NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHORS
PHOSPHORUS ISOTOPES
POLYMERIZATION
RADIOISOTOPES
SODIUM COMPOUNDS
550201* - Biochemistry- Tracer Techniques