Myeloperoxidase-Catalyzed Incorporation of Amines into Proteins: Role of Hypochlorous Acid and Dichloramines

Edwin Thomas, M. Margaret Jefferson, Matthew B. Grisham, Edwin L. Thomas, M. Margaret Jefferson, Matthew B. Grisham

Research output: Contribution to journalArticle

103 Citations (Scopus)

Abstract

Myeloperoxidase-catalyzed oxidation of chloride (Cl) to hypochlorous acid (HOC1) resulted in formation of mono- and dichloramine derivatives (RNHC1 and RNC12) of primary amines. The RNC12 derivatives could undergo a reaction that resulted in incorporation of the R moiety into proteins. The probable mechanism was attack of RNC12 or an intermediate formed in the decomposition of RNC12 on histidine, tyrosine, and cystine residues and on lysine residues at high pH. Incorporation of radioactivity from labeled amines into stable, high molecular weight derivatives of proteins was measured by acid or acetone precipitation and by gel chromatography and electrophoresis. Whereas formation of RNCl2was favored at low pH, the subsequent incorporation reaction was favored at high pH. Up to several hours were required for the maximum amount of incorporation, which was less than 10% of the label in RNCl2. For the amines tested, incorporation was in the order histamine > 1,2-diaminoethane > putrescine > taurine > lysine > glucosamine > leucine > methylamine. Initiation of the reaction required HOC1, and oxidized forms of bromide, iodide, or thiocyanate did not substitute. Inhibitors of incorporation fell into three classes. First, ammonia or amines competed with the labeled amine for reaction with HOC1, so that larger amounts of HOC1 were required. Second, readily oxidized substances such as sulfhydryl or diketo compounds or thioethers (methionine) reduced RNC12. Third, certain compounds competed with protein as the acceptor for the incorporation reaction. The amount required to block incorporation into protein depended on protein concentration. Among these inhibitors were imidazole compounds (histidine), phenols (tyrosine), and disulfides (glutathione disulfide, GSSG). Low yields of derivatives of histidine, tyrosine, and GSSG were detected by thin-layer chromatography. Acid-precipitable derivatives were obtained by reacting RNCl2 with polyhistidine or polytyrosine, and to a lesser extent with polylysine at high pH, but not with other poly(amino acids). Precipitable derivatives were also obtained by incubating MPO-containing extracts from leukocyte granules with hydrogen peroxide, Cl, and labeled amines. The extracts were found to have a high content of substances with primary amino groups, which competed for incorporation. The results account for oxidative incorporation of amines into proteins in leukocytes and provide evidence that HOCl and nitrogen-chlorine (N-Cl) derivatives are formed in these cells. The characteristics of the incorporation reaction suggest that it would not contribute significantly to the antimicrobial activity of myeloperoxidase (MPO). Nevertheless, the reaction may provide a sensitive method for studying MPO action in vivo.

Original languageEnglish (US)
Pages (from-to)6299-6308
Number of pages10
JournalBiochemistry
Volume21
Issue number24
DOIs
StatePublished - Jan 1 1982

Fingerprint

Hypochlorous Acid
Peroxidase
Amines
Derivatives
Glutathione Disulfide
Histidine
Proteins
Tyrosine
ethylenediamine
Lysine
Chlorides
Leukocytes
Thin layer chromatography
Acids
Polylysine
Putrescine
Cystine
Hydrochloric Acid
Phenols
Taurine

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Thomas, E., Jefferson, M. M., Grisham, M. B., Thomas, E. L., Jefferson, M. M., & Grisham, M. B. (1982). Myeloperoxidase-Catalyzed Incorporation of Amines into Proteins: Role of Hypochlorous Acid and Dichloramines. Biochemistry, 21(24), 6299-6308. https://doi.org/10.1021/bi00267a040

Myeloperoxidase-Catalyzed Incorporation of Amines into Proteins : Role of Hypochlorous Acid and Dichloramines. / Thomas, Edwin; Jefferson, M. Margaret; Grisham, Matthew B.; Thomas, Edwin L.; Jefferson, M. Margaret; Grisham, Matthew B.

In: Biochemistry, Vol. 21, No. 24, 01.01.1982, p. 6299-6308.

Research output: Contribution to journalArticle

Thomas, E, Jefferson, MM, Grisham, MB, Thomas, EL, Jefferson, MM & Grisham, MB 1982, 'Myeloperoxidase-Catalyzed Incorporation of Amines into Proteins: Role of Hypochlorous Acid and Dichloramines', Biochemistry, vol. 21, no. 24, pp. 6299-6308. https://doi.org/10.1021/bi00267a040
Thomas, Edwin ; Jefferson, M. Margaret ; Grisham, Matthew B. ; Thomas, Edwin L. ; Jefferson, M. Margaret ; Grisham, Matthew B. / Myeloperoxidase-Catalyzed Incorporation of Amines into Proteins : Role of Hypochlorous Acid and Dichloramines. In: Biochemistry. 1982 ; Vol. 21, No. 24. pp. 6299-6308.
@article{ee1b37539ae44b5aa9e44e67a9557e3a,
title = "Myeloperoxidase-Catalyzed Incorporation of Amines into Proteins: Role of Hypochlorous Acid and Dichloramines",
abstract = "Myeloperoxidase-catalyzed oxidation of chloride (Cl−) to hypochlorous acid (HOC1) resulted in formation of mono- and dichloramine derivatives (RNHC1 and RNC12) of primary amines. The RNC12 derivatives could undergo a reaction that resulted in incorporation of the R moiety into proteins. The probable mechanism was attack of RNC12 or an intermediate formed in the decomposition of RNC12 on histidine, tyrosine, and cystine residues and on lysine residues at high pH. Incorporation of radioactivity from labeled amines into stable, high molecular weight derivatives of proteins was measured by acid or acetone precipitation and by gel chromatography and electrophoresis. Whereas formation of RNCl2was favored at low pH, the subsequent incorporation reaction was favored at high pH. Up to several hours were required for the maximum amount of incorporation, which was less than 10{\%} of the label in RNCl2. For the amines tested, incorporation was in the order histamine > 1,2-diaminoethane > putrescine > taurine > lysine > glucosamine > leucine > methylamine. Initiation of the reaction required HOC1, and oxidized forms of bromide, iodide, or thiocyanate did not substitute. Inhibitors of incorporation fell into three classes. First, ammonia or amines competed with the labeled amine for reaction with HOC1, so that larger amounts of HOC1 were required. Second, readily oxidized substances such as sulfhydryl or diketo compounds or thioethers (methionine) reduced RNC12. Third, certain compounds competed with protein as the acceptor for the incorporation reaction. The amount required to block incorporation into protein depended on protein concentration. Among these inhibitors were imidazole compounds (histidine), phenols (tyrosine), and disulfides (glutathione disulfide, GSSG). Low yields of derivatives of histidine, tyrosine, and GSSG were detected by thin-layer chromatography. Acid-precipitable derivatives were obtained by reacting RNCl2 with polyhistidine or polytyrosine, and to a lesser extent with polylysine at high pH, but not with other poly(amino acids). Precipitable derivatives were also obtained by incubating MPO-containing extracts from leukocyte granules with hydrogen peroxide, Cl−, and labeled amines. The extracts were found to have a high content of substances with primary amino groups, which competed for incorporation. The results account for oxidative incorporation of amines into proteins in leukocytes and provide evidence that HOCl and nitrogen-chlorine (N-Cl) derivatives are formed in these cells. The characteristics of the incorporation reaction suggest that it would not contribute significantly to the antimicrobial activity of myeloperoxidase (MPO). Nevertheless, the reaction may provide a sensitive method for studying MPO action in vivo.",
author = "Edwin Thomas and Jefferson, {M. Margaret} and Grisham, {Matthew B.} and Thomas, {Edwin L.} and Jefferson, {M. Margaret} and Grisham, {Matthew B.}",
year = "1982",
month = "1",
day = "1",
doi = "10.1021/bi00267a040",
language = "English (US)",
volume = "21",
pages = "6299--6308",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "24",

}

TY - JOUR

T1 - Myeloperoxidase-Catalyzed Incorporation of Amines into Proteins

T2 - Role of Hypochlorous Acid and Dichloramines

AU - Thomas, Edwin

AU - Jefferson, M. Margaret

AU - Grisham, Matthew B.

AU - Thomas, Edwin L.

AU - Jefferson, M. Margaret

AU - Grisham, Matthew B.

PY - 1982/1/1

Y1 - 1982/1/1

N2 - Myeloperoxidase-catalyzed oxidation of chloride (Cl−) to hypochlorous acid (HOC1) resulted in formation of mono- and dichloramine derivatives (RNHC1 and RNC12) of primary amines. The RNC12 derivatives could undergo a reaction that resulted in incorporation of the R moiety into proteins. The probable mechanism was attack of RNC12 or an intermediate formed in the decomposition of RNC12 on histidine, tyrosine, and cystine residues and on lysine residues at high pH. Incorporation of radioactivity from labeled amines into stable, high molecular weight derivatives of proteins was measured by acid or acetone precipitation and by gel chromatography and electrophoresis. Whereas formation of RNCl2was favored at low pH, the subsequent incorporation reaction was favored at high pH. Up to several hours were required for the maximum amount of incorporation, which was less than 10% of the label in RNCl2. For the amines tested, incorporation was in the order histamine > 1,2-diaminoethane > putrescine > taurine > lysine > glucosamine > leucine > methylamine. Initiation of the reaction required HOC1, and oxidized forms of bromide, iodide, or thiocyanate did not substitute. Inhibitors of incorporation fell into three classes. First, ammonia or amines competed with the labeled amine for reaction with HOC1, so that larger amounts of HOC1 were required. Second, readily oxidized substances such as sulfhydryl or diketo compounds or thioethers (methionine) reduced RNC12. Third, certain compounds competed with protein as the acceptor for the incorporation reaction. The amount required to block incorporation into protein depended on protein concentration. Among these inhibitors were imidazole compounds (histidine), phenols (tyrosine), and disulfides (glutathione disulfide, GSSG). Low yields of derivatives of histidine, tyrosine, and GSSG were detected by thin-layer chromatography. Acid-precipitable derivatives were obtained by reacting RNCl2 with polyhistidine or polytyrosine, and to a lesser extent with polylysine at high pH, but not with other poly(amino acids). Precipitable derivatives were also obtained by incubating MPO-containing extracts from leukocyte granules with hydrogen peroxide, Cl−, and labeled amines. The extracts were found to have a high content of substances with primary amino groups, which competed for incorporation. The results account for oxidative incorporation of amines into proteins in leukocytes and provide evidence that HOCl and nitrogen-chlorine (N-Cl) derivatives are formed in these cells. The characteristics of the incorporation reaction suggest that it would not contribute significantly to the antimicrobial activity of myeloperoxidase (MPO). Nevertheless, the reaction may provide a sensitive method for studying MPO action in vivo.

AB - Myeloperoxidase-catalyzed oxidation of chloride (Cl−) to hypochlorous acid (HOC1) resulted in formation of mono- and dichloramine derivatives (RNHC1 and RNC12) of primary amines. The RNC12 derivatives could undergo a reaction that resulted in incorporation of the R moiety into proteins. The probable mechanism was attack of RNC12 or an intermediate formed in the decomposition of RNC12 on histidine, tyrosine, and cystine residues and on lysine residues at high pH. Incorporation of radioactivity from labeled amines into stable, high molecular weight derivatives of proteins was measured by acid or acetone precipitation and by gel chromatography and electrophoresis. Whereas formation of RNCl2was favored at low pH, the subsequent incorporation reaction was favored at high pH. Up to several hours were required for the maximum amount of incorporation, which was less than 10% of the label in RNCl2. For the amines tested, incorporation was in the order histamine > 1,2-diaminoethane > putrescine > taurine > lysine > glucosamine > leucine > methylamine. Initiation of the reaction required HOC1, and oxidized forms of bromide, iodide, or thiocyanate did not substitute. Inhibitors of incorporation fell into three classes. First, ammonia or amines competed with the labeled amine for reaction with HOC1, so that larger amounts of HOC1 were required. Second, readily oxidized substances such as sulfhydryl or diketo compounds or thioethers (methionine) reduced RNC12. Third, certain compounds competed with protein as the acceptor for the incorporation reaction. The amount required to block incorporation into protein depended on protein concentration. Among these inhibitors were imidazole compounds (histidine), phenols (tyrosine), and disulfides (glutathione disulfide, GSSG). Low yields of derivatives of histidine, tyrosine, and GSSG were detected by thin-layer chromatography. Acid-precipitable derivatives were obtained by reacting RNCl2 with polyhistidine or polytyrosine, and to a lesser extent with polylysine at high pH, but not with other poly(amino acids). Precipitable derivatives were also obtained by incubating MPO-containing extracts from leukocyte granules with hydrogen peroxide, Cl−, and labeled amines. The extracts were found to have a high content of substances with primary amino groups, which competed for incorporation. The results account for oxidative incorporation of amines into proteins in leukocytes and provide evidence that HOCl and nitrogen-chlorine (N-Cl) derivatives are formed in these cells. The characteristics of the incorporation reaction suggest that it would not contribute significantly to the antimicrobial activity of myeloperoxidase (MPO). Nevertheless, the reaction may provide a sensitive method for studying MPO action in vivo.

UR - http://www.scopus.com/inward/record.url?scp=0020361724&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020361724&partnerID=8YFLogxK

U2 - 10.1021/bi00267a040

DO - 10.1021/bi00267a040

M3 - Article

C2 - 6295461

AN - SCOPUS:0020361724

VL - 21

SP - 6299

EP - 6308

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 24

ER -