Development of a novel photoreactive calmodulin derivative: Cross-linking of purified adenylate cyclase from bovine brain

Journal Article · Tue Jul 11 04:00:00 EDT 1989 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00440a045· OSTI ID:7018934

Harrison, J K; Lawton, R G; Gnegy, M E ^[1]

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1. 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), Journal Name: Biochemistry; (USA) Vol. 28:14; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALI METAL COMPOUNDS
ALKALINE EARTH METAL COMPOUNDS
ATP
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
BRAIN
CALCIUM CHLORIDES
CALCIUM COMPOUNDS
CALCIUM HALIDES
CALMODULIN
CENTRAL NERVOUS SYSTEM
CHEMICAL REACTIONS
CHLORIDES
CHLORINE COMPOUNDS
CROSS-LINKING
CYCLASES
DAYS LIVING RADIOISOTOPES
DERIVATIZATION
ELECTRON CAPTURE RADIOISOTOPES
ENZYME ACTIVITY
ENZYMES
HALIDES
HALOGEN COMPOUNDS
HYDROGEN COMPOUNDS
INORGANIC PHOSPHORS
INTERMEDIATE MASS NUCLEI
IODIDES
IODINE 125
IODINE COMPOUNDS
IODINE ISOTOPES
ISOTOPES
LIGHT NUCLEI
LYASES
NERVOUS SYSTEM
NUCLEI
NUCLEOTIDES
ODD-EVEN NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHORS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
POLYMERIZATION
PROTEINS
RADIOISOTOPES
SODIUM COMPOUNDS
SODIUM IODIDES
TRITIUM COMPOUNDS