Biotinylated human. beta. -endorphins as probes for the opioid receptor

Hochhaus, G; Gibson, B W; Sadee, W

Title: Biotinylated human. beta. -endorphins as probes for the opioid receptor

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Abstract

The reaction of human ..beta..-endorphin and biotinyl N-hydroxysuccinimide with or without spacer arm, afforded a series of products that were separated by high performance liquid chromatography (HPLC). Liquid secondary ion mass spectrometry of the biotinylated products and their tryptic digests produced abundant protonated molecular ions (MH/sup +/), which specified the number and location of biotinylation. Between 1 and 4 biotinyl residues were incorporated per human ..beta..-endorphin molecule, at Lys-9, -19, -24, -28, and -29, but not at the amino-terminal Try-1. Three HPLC fractions were isolated for receptor binding studies monobiotinylation of Lys-9, Lys-19, and a mixture of Lys-24, Lys-28, and Lys-29 derivatives. IC/sub 50/ values for binding to ..mu.. and delta opioid receptor sites were 3-8 times higher for monobiotinylated derivatives than for the parent human ..beta..-endorphin. Association with avidin decreased opioid receptor affinities for the C/sub 6/ spacer derivative biotinylated at position Lys-9, which is close to the (1-5) enkephalin receptor region. In contrast, avidin did not affect or even increased apparent affinities to ..mu.. and delta sites for derivatives biotinylated at the ..cap alpha..-helical part of the molecule (Lys-19, -24, -28, and -29). Biotinylated human ..beta..-endorphins also bound to low affinity nonopioid binding sites on NG-108-15 cells; however,more »« less

Authors:: Hochhaus, G; Gibson, B W; Sadee, W

Publication Date:: Tue Jan 05 00:00:00 EST 1988

Research Org.:: Univ. of California, San Francisco (USA)

OSTI Identifier:: 6922187

Resource Type:: Journal Article

Journal Name:: J. Biol. Chem.; (United States)

Additional Journal Information:: Journal Volume: 263:1

Country of Publication:: United States

Language:: English

Subject:: 63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; 3-METHYLCHOLANTHRENE; BIOLOGICAL EFFECTS; PHENOBARBITAL; TRIPHENYLENE; METABOLISM; CARCINOGENESIS; HYDROLASES; LIVER; MICROSOMES; RACEMIZATION; RATS; ANESTHETICS; ANIMALS; ANTICONVULSANTS; AROMATICS; AZINES; BARBITURATES; BODY; CELL CONSTITUENTS; CENTRAL NERVOUS SYSTEM DEPRESSANTS; CONDENSED AROMATICS; DIGESTIVE SYSTEM; DRUGS; ENZYMES; GLANDS; HETEROCYCLIC COMPOUNDS; HYDROCARBONS; HYPNOTICS AND SEDATIVES; MAMMALS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORGANIC OXYGEN COMPOUNDS; ORGANOIDS; ORGANS; PATHOGENESIS; POLYCYCLIC AROMATIC HYDROCARBONS; PYRIMIDINES; RODENTS; VERTEBRATES; 560300* - Chemicals Metabolism & Toxicology

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                    Hochhaus, G, Gibson, B W, and Sadee, W. Biotinylated human. beta. -endorphins as probes for the opioid receptor.  United States: N. p., 1988. 
        Web.

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                    Hochhaus, G, Gibson, B W, & Sadee, W. Biotinylated human. beta. -endorphins as probes for the opioid receptor.  United States.

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                    Hochhaus, G, Gibson, B W, and Sadee, W. 1988.  
        "Biotinylated human. beta. -endorphins as probes for the opioid receptor".  United States.

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@article{osti_6922187,

  title        = {Biotinylated human. beta. -endorphins as probes for the opioid receptor},

  author       = {Hochhaus, G and Gibson, B W and Sadee, W},

  abstractNote = {The reaction of human ..beta..-endorphin and biotinyl N-hydroxysuccinimide with or without spacer arm, afforded a series of products that were separated by high performance liquid chromatography (HPLC). Liquid secondary ion mass spectrometry of the biotinylated products and their tryptic digests produced abundant protonated molecular ions (MH/sup +/), which specified the number and location of biotinylation. Between 1 and 4 biotinyl residues were incorporated per human ..beta..-endorphin molecule, at Lys-9, -19, -24, -28, and -29, but not at the amino-terminal Try-1. Three HPLC fractions were isolated for receptor binding studies monobiotinylation of Lys-9, Lys-19, and a mixture of Lys-24, Lys-28, and Lys-29 derivatives. IC/sub 50/ values for binding to ..mu.. and delta opioid receptor sites were 3-8 times higher for monobiotinylated derivatives than for the parent human ..beta..-endorphin. Association with avidin decreased opioid receptor affinities for the C/sub 6/ spacer derivative biotinylated at position Lys-9, which is close to the (1-5) enkephalin receptor region. In contrast, avidin did not affect or even increased apparent affinities to ..mu.. and delta sites for derivatives biotinylated at the ..cap alpha..-helical part of the molecule (Lys-19, -24, -28, and -29). Biotinylated human ..beta..-endorphins also bound to low affinity nonopioid binding sites on NG-108-15 cells; however, affinities to these sites were considerably reduced when derivatives were bound to avidin. The ability of biotinylated human ..beta..-endorphin to cross-link the ..mu.. and delta opioid receptors to avidin allows application of the biotin-avidin system as a molecular probe of the opioid receptor.},

  doi          = {},

  url          = {https://www.osti.gov/biblio/6922187},
  journal      = {J. Biol. Chem.; (United States)},
number       = ,

  volume       = 263:1,

  place        = {United States},

  year         = {Tue Jan 05 00:00:00 EST 1988},

  month        = {Tue Jan 05 00:00:00 EST 1988}

}

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