Determination of Tissue Histamine by an Enzymatic Isotopic Assya

Solomon H. Snyder

doi:10.1016/0076-6879(71)17149-2

Determination of Tissue Histamine by an Enzymatic Isotopic Assya

Solomon H. Snyder

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Abstract

This chapter discusses determination of tissue histamine by an enzymatic isotopic assay. The method depends on the enzymatic transfer of the methyl-¹⁴C of S-adenosylmethionine-¹⁴C (AMe-¹⁴C) to tissue histamine to form methylhistamine-¹⁴C. A tracer amount of histamine-all is added to endogenous histamine; the percent of the tritium in the methylhistamine formed indicates the extent of the enzymatic methylation. The final product formed is methylhistamine-¹⁴C-³H. Endogenous AMe is destroyed by heating so that there is no dilution of the added ¹⁴C-AMe-¹⁴C. The enzyme histamine methyl transferase used for the reaction is highly specific for histamine. The only tissue component that is methylated in the reaction is histamine; chloroform extraction separates histamine from methylhistamine. Thus the ratio ¹⁴C:³H is linearly related to the tissue content of histamine. With each assay are included standard determinations of two concentrations of authentic histamine as well as histamine added to tissue homogenates before heating. Histamine added to tissues is recovered to the extent of 90-100%.

Original language	English (US)
Pages (from-to)	838-841
Number of pages	4
Journal	Methods in enzymology
Volume	17
DOIs	https://doi.org/10.1016/0076-6879(71)17149-2
State	Published - Jan 1 1971
Externally published	Yes

ASJC Scopus subject areas

Biochemistry
Molecular Biology

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10.1016/0076-6879(71)17149-2

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title = "Determination of Tissue Histamine by an Enzymatic Isotopic Assya",

abstract = "This chapter discusses determination of tissue histamine by an enzymatic isotopic assay. The method depends on the enzymatic transfer of the methyl-14C of S-adenosylmethionine-14C (AMe-14C) to tissue histamine to form methylhistamine-14C. A tracer amount of histamine-all is added to endogenous histamine; the percent of the tritium in the methylhistamine formed indicates the extent of the enzymatic methylation. The final product formed is methylhistamine-14C-3H. Endogenous AMe is destroyed by heating so that there is no dilution of the added 14C-AMe-14C. The enzyme histamine methyl transferase used for the reaction is highly specific for histamine. The only tissue component that is methylated in the reaction is histamine; chloroform extraction separates histamine from methylhistamine. Thus the ratio 14C:3H is linearly related to the tissue content of histamine. With each assay are included standard determinations of two concentrations of authentic histamine as well as histamine added to tissue homogenates before heating. Histamine added to tissues is recovered to the extent of 90-100%.",

author = "Snyder, {Solomon H.}",

note = "Funding Information: Histamine-3H generally labeled (9.6 C/millimole) S-Adenosylmethionine-14C (40 mC/millimole) Histamine methyl transferase is partially purified by a modification of the method of Brown et al. 3 The brains of 6 adult male guinea pigs 1This work was supported, in part, by NIH grants 1-MH-11267 and 1-RO1-NB-07275. The author is a recipient of NIMH Research Career Development Award K-3-MH-33128. 2D. D. Brown, R. Tomchick, and J. Axelrod, J. Biol. Chem. 234, 2948 (1959). Histamine N-methyhransferase has also been purified 300-fold by Gustafsson and Forschell \[Acta Chem. Scand. 17,541 (1963)\]f rom hog liver. For a more highly purified preparation of the brain enzyme, seeJ. Axelrod, this volume \[243c\]. 3S. H. Snyder, R.J. Baldessarini, and J. Axelrod,J. Pharm. Exptl. Therap. 153, 544 (1966).",

year = "1971",

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doi = "10.1016/0076-6879(71)17149-2",

language = "English (US)",

volume = "17",

pages = "838--841",

journal = "Methods in enzymology",

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T1 - Determination of Tissue Histamine by an Enzymatic Isotopic Assya

AU - Snyder, Solomon H.

N1 - Funding Information: Histamine-3H generally labeled (9.6 C/millimole) S-Adenosylmethionine-14C (40 mC/millimole) Histamine methyl transferase is partially purified by a modification of the method of Brown et al. 3 The brains of 6 adult male guinea pigs 1This work was supported, in part, by NIH grants 1-MH-11267 and 1-RO1-NB-07275. The author is a recipient of NIMH Research Career Development Award K-3-MH-33128. 2D. D. Brown, R. Tomchick, and J. Axelrod, J. Biol. Chem. 234, 2948 (1959). Histamine N-methyhransferase has also been purified 300-fold by Gustafsson and Forschell \[Acta Chem. Scand. 17,541 (1963)\]f rom hog liver. For a more highly purified preparation of the brain enzyme, seeJ. Axelrod, this volume \[243c\]. 3S. H. Snyder, R.J. Baldessarini, and J. Axelrod,J. Pharm. Exptl. Therap. 153, 544 (1966).

PY - 1971/1/1

Y1 - 1971/1/1

N2 - This chapter discusses determination of tissue histamine by an enzymatic isotopic assay. The method depends on the enzymatic transfer of the methyl-14C of S-adenosylmethionine-14C (AMe-14C) to tissue histamine to form methylhistamine-14C. A tracer amount of histamine-all is added to endogenous histamine; the percent of the tritium in the methylhistamine formed indicates the extent of the enzymatic methylation. The final product formed is methylhistamine-14C-3H. Endogenous AMe is destroyed by heating so that there is no dilution of the added 14C-AMe-14C. The enzyme histamine methyl transferase used for the reaction is highly specific for histamine. The only tissue component that is methylated in the reaction is histamine; chloroform extraction separates histamine from methylhistamine. Thus the ratio 14C:3H is linearly related to the tissue content of histamine. With each assay are included standard determinations of two concentrations of authentic histamine as well as histamine added to tissue homogenates before heating. Histamine added to tissues is recovered to the extent of 90-100%.

AB - This chapter discusses determination of tissue histamine by an enzymatic isotopic assay. The method depends on the enzymatic transfer of the methyl-14C of S-adenosylmethionine-14C (AMe-14C) to tissue histamine to form methylhistamine-14C. A tracer amount of histamine-all is added to endogenous histamine; the percent of the tritium in the methylhistamine formed indicates the extent of the enzymatic methylation. The final product formed is methylhistamine-14C-3H. Endogenous AMe is destroyed by heating so that there is no dilution of the added 14C-AMe-14C. The enzyme histamine methyl transferase used for the reaction is highly specific for histamine. The only tissue component that is methylated in the reaction is histamine; chloroform extraction separates histamine from methylhistamine. Thus the ratio 14C:3H is linearly related to the tissue content of histamine. With each assay are included standard determinations of two concentrations of authentic histamine as well as histamine added to tissue homogenates before heating. Histamine added to tissues is recovered to the extent of 90-100%.

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DO - 10.1016/0076-6879(71)17149-2

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JO - Methods in enzymology

JF - Methods in enzymology

ER -

Determination of Tissue Histamine by an Enzymatic Isotopic Assya

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