Effect of buffer composition on binding of adenosine 3':5'-cyclic monophosphate by bovine adrenal extracts

The binding of cyclic [3H] AMP by bovine adrenal extracts was studied in different buffer systems; 50 mM-Tris/HCl, pH 7.5, buffer containing 4mM-EDTA (disodium salt) offered advantages such as low non-specific interference and high binding affinity. 2-Mercaptoethanol decreased, but EDTA and theophyl...

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Veröffentlicht in:	Biochemical journal 1977-09, Vol.166 (3), p.553-557
Hauptverfasser:	Linkola, J, Fyhrquist, F, Weber, T H, Puutula, L
Format:	Artikel
Sprache:	eng
Schlagworte:	Adrenal Glands - drug effects Adrenal Glands - metabolism Buffers Cyclic AMP - metabolism Edetic Acid - pharmacology In Vitro Techniques Magnesium Mercaptoethanol - pharmacology Methods Theophylline - pharmacology Tissue Extracts - metabolism
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container_title	Biochemical journal
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description	The binding of cyclic [3H] AMP by bovine adrenal extracts was studied in different buffer systems; 50 mM-Tris/HCl, pH 7.5, buffer containing 4mM-EDTA (disodium salt) offered advantages such as low non-specific interference and high binding affinity. 2-Mercaptoethanol decreased, but EDTA and theophylline increased, the specificity of cyclic [3H]AMP binding as expressed by the displacement of cyclic [3H]AMP by a 1000-fold excess of unlabelled cyclic GMP. The study indicated heterogeneity of the binding material. Mg2+-dependent binding may be eliminated in Tris/EDTA buffer. The heterogeneity and the specificity of intracellular cyclic AMP binders and their dependence in environmental factors should be considered in the evaluation of cyclic AMP binding and second-messenger systems.
doi_str_mv	10.1042/bj1660553
format	Article
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The study indicated heterogeneity of the binding material. Mg2+-dependent binding may be eliminated in Tris/EDTA buffer. 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The study indicated heterogeneity of the binding material. Mg2+-dependent binding may be eliminated in Tris/EDTA buffer. The heterogeneity and the specificity of intracellular cyclic AMP binders and their dependence in environmental factors should be considered in the evaluation of cyclic AMP binding and second-messenger systems.</description><subject>Adrenal Glands - drug effects</subject><subject>Adrenal Glands - metabolism</subject><subject>Buffers</subject><subject>Cyclic AMP - metabolism</subject><subject>Edetic Acid - pharmacology</subject><subject>In Vitro Techniques</subject><subject>Magnesium</subject><subject>Mercaptoethanol - pharmacology</subject><subject>Methods</subject><subject>Theophylline - pharmacology</subject><subject>Tissue Extracts - metabolism</subject><issn>0264-6021</issn><issn>1470-8728</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1977</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkctLAzEQxoP4qtWDdw85WTys5rGbtB4EKb6g4EXPIcnO2pTdZN3siv3vTakUhYEZ-H58M8yH0Dkl15Tk7MasqBCkKPgeGtFckmwq2XQfjQgTeSYIo8foJMYVITQnOTlCh4wwls9GaPVQVWB7HCpshjR22IamDdH1LnicyjhfOv-xAXQJPikeMJ_cFpPMrm3tLG6CD-0yxHape8BmjU342kC67MDrGsN332nbx1N0UOk6wtlvH6P3x4e3-XO2eH16md8vMsvFrM8Y4VxqKjSVFeV5IZngwDhjYCpKSsElNRWZTTkFzYw1xVSAkZyaUoAujeBjdLf1bQfTQGnBp_21ajvX6G6tgnbqv-LdUn2EL0WpKNJ3ksHlr0EXPgeIvWpctFDX2kMYopJcyvRfnsCrLWi7EGMH1W4JJWqTi9rlktiLv1ftyG0Q_AfiCYmA</recordid><startdate>19770915</startdate><enddate>19770915</enddate><creator>Linkola, J</creator><creator>Fyhrquist, F</creator><creator>Weber, T H</creator><creator>Puutula, L</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19770915</creationdate><title>Effect of buffer composition on binding of adenosine 3':5'-cyclic monophosphate by bovine adrenal extracts</title><author>Linkola, J ; Fyhrquist, F ; Weber, T H ; Puutula, L</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c369t-20337a16a17f13457263e2322ebf10d6371bf09831ea2bcb586eb731bd6eadb63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1977</creationdate><topic>Adrenal Glands - drug effects</topic><topic>Adrenal Glands - metabolism</topic><topic>Buffers</topic><topic>Cyclic AMP - metabolism</topic><topic>Edetic Acid - pharmacology</topic><topic>In Vitro Techniques</topic><topic>Magnesium</topic><topic>Mercaptoethanol - pharmacology</topic><topic>Methods</topic><topic>Theophylline - pharmacology</topic><topic>Tissue Extracts - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Linkola, J</creatorcontrib><creatorcontrib>Fyhrquist, F</creatorcontrib><creatorcontrib>Weber, T H</creatorcontrib><creatorcontrib>Puutula, L</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Biochemical journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Linkola, J</au><au>Fyhrquist, F</au><au>Weber, T H</au><au>Puutula, L</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effect of buffer composition on binding of adenosine 3':5'-cyclic monophosphate by bovine adrenal extracts</atitle><jtitle>Biochemical journal</jtitle><addtitle>Biochem J</addtitle><date>1977-09-15</date><risdate>1977</risdate><volume>166</volume><issue>3</issue><spage>553</spage><epage>557</epage><pages>553-557</pages><issn>0264-6021</issn><eissn>1470-8728</eissn><abstract>The binding of cyclic [3H] AMP by bovine adrenal extracts was studied in different buffer systems; 50 mM-Tris/HCl, pH 7.5, buffer containing 4mM-EDTA (disodium salt) offered advantages such as low non-specific interference and high binding affinity. 2-Mercaptoethanol decreased, but EDTA and theophylline increased, the specificity of cyclic [3H]AMP binding as expressed by the displacement of cyclic [3H]AMP by a 1000-fold excess of unlabelled cyclic GMP. 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subjects	Adrenal Glands - drug effects Adrenal Glands - metabolism Buffers Cyclic AMP - metabolism Edetic Acid - pharmacology In Vitro Techniques Magnesium Mercaptoethanol - pharmacology Methods Theophylline - pharmacology Tissue Extracts - metabolism
title	Effect of buffer composition on binding of adenosine 3':5'-cyclic monophosphate by bovine adrenal extracts
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