Evidence for inactivation of cysteine proteases by reactive carbonyls via glycation of active site thiols

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

Evidence for inactivation of cysteine proteases by reactive carbonyls via glycation of active site thiols. / Zeng, Jingmin; Dunlop, Rachael A; Rodgers, Kenneth J; Davies, Michael Jonathan.

In: Biochemical Journal, Vol. 398, No. 2, 01.09.2006, p. 197-206.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Zeng, J, Dunlop, RA, Rodgers, KJ & Davies, MJ 2006, 'Evidence for inactivation of cysteine proteases by reactive carbonyls via glycation of active site thiols', Biochemical Journal, vol. 398, no. 2, pp. 197-206. https://doi.org/10.1042/BJ20060019

APA

Zeng, J., Dunlop, R. A., Rodgers, K. J., & Davies, M. J. (2006). Evidence for inactivation of cysteine proteases by reactive carbonyls via glycation of active site thiols. Biochemical Journal, 398(2), 197-206. https://doi.org/10.1042/BJ20060019

Vancouver

Zeng J, Dunlop RA, Rodgers KJ, Davies MJ. Evidence for inactivation of cysteine proteases by reactive carbonyls via glycation of active site thiols. Biochemical Journal. 2006 Sep 1;398(2):197-206. https://doi.org/10.1042/BJ20060019

Author

Zeng, Jingmin ; Dunlop, Rachael A ; Rodgers, Kenneth J ; Davies, Michael Jonathan. / Evidence for inactivation of cysteine proteases by reactive carbonyls via glycation of active site thiols. In: Biochemical Journal. 2006 ; Vol. 398, No. 2. pp. 197-206.

Bibtex

@article{f985d8673b8046e09941e8082e5adb85,
title = "Evidence for inactivation of cysteine proteases by reactive carbonyls via glycation of active site thiols",
abstract = "Hyperglycaemia, triose phosphate decomposition and oxidation reactions generate reactive aldehydes in vivo. These compounds react non-enzymatically with protein side chains and N-terminal amino groups to give adducts and cross-links, and hence modified proteins. Previous studies have shown that free or protein-bound carbonyls inactivate glyceraldehyde-3-phosphate dehydrogenase with concomitant loss of thiol groups [Morgan, Dean and Davies (2002) Arch. Biochem. Biophys. 403, 259-269]. It was therefore hypothesized that modification of lysosomal cysteine proteases (and the structurally related enzyme papain) by free and protein-bound carbonyls may modulate the activity of these components of the cellular proteolytic machinery responsible for the removal of modified proteins and thereby contribute to a decreased removal of modified proteins from cells. It is shown that MGX (methylglyoxal), GO (glyoxal) and glycolaldehyde, but not hydroxyacetone and glucose, inhibit catB (cathepsin B), catL (cathepsin L) and catS (cathepsin S) activity in macrophage cell lysates, in a concentration-dependent manner. Protein-bound carbonyls produced similar inhibition with both cell lysates and intact macrophage cells. Inhibition was also observed with papain, with this paralleled by loss of the active site cysteine residue and formation of the adduct species S-carboxymethylcysteine, from GO, in a concentration-dependent manner. Inhibition of autolysis of papain by MGX, along with cross-link formation, was detected by SDS/PAGE. Treatment of papain and catS with the dialdehyde o-phthalaldehyde resulted in enzyme inactivation and an intra-molecular active site cysteine-lysine cross-link. These results demonstrate that reactive aldehydes inhibit cysteine proteases by modification of the active site cysteine residue. This process may contribute to the accumulation of modified proteins in tissues of people with diabetes and age-related pathologies, including atherosclerosis, cataract and Alzheimer's disease.",
keywords = "Animals, Binding Sites, Carbocysteine, Cathepsins, Cattle, Cell Line, Cysteine Endopeptidases, Enzyme Activation, Glycosylation, Glyoxal, Humans, Mice, Molecular Weight, Papain, Protein Binding, Sulfhydryl Compounds, o-Phthalaldehyde",
author = "Jingmin Zeng and Dunlop, {Rachael A} and Rodgers, {Kenneth J} and Davies, {Michael Jonathan}",
year = "2006",
month = "9",
day = "1",
doi = "10.1042/BJ20060019",
language = "English",
volume = "398",
pages = "197--206",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

RIS

TY - JOUR

T1 - Evidence for inactivation of cysteine proteases by reactive carbonyls via glycation of active site thiols

AU - Zeng, Jingmin

AU - Dunlop, Rachael A

AU - Rodgers, Kenneth J

AU - Davies, Michael Jonathan

PY - 2006/9/1

Y1 - 2006/9/1

N2 - Hyperglycaemia, triose phosphate decomposition and oxidation reactions generate reactive aldehydes in vivo. These compounds react non-enzymatically with protein side chains and N-terminal amino groups to give adducts and cross-links, and hence modified proteins. Previous studies have shown that free or protein-bound carbonyls inactivate glyceraldehyde-3-phosphate dehydrogenase with concomitant loss of thiol groups [Morgan, Dean and Davies (2002) Arch. Biochem. Biophys. 403, 259-269]. It was therefore hypothesized that modification of lysosomal cysteine proteases (and the structurally related enzyme papain) by free and protein-bound carbonyls may modulate the activity of these components of the cellular proteolytic machinery responsible for the removal of modified proteins and thereby contribute to a decreased removal of modified proteins from cells. It is shown that MGX (methylglyoxal), GO (glyoxal) and glycolaldehyde, but not hydroxyacetone and glucose, inhibit catB (cathepsin B), catL (cathepsin L) and catS (cathepsin S) activity in macrophage cell lysates, in a concentration-dependent manner. Protein-bound carbonyls produced similar inhibition with both cell lysates and intact macrophage cells. Inhibition was also observed with papain, with this paralleled by loss of the active site cysteine residue and formation of the adduct species S-carboxymethylcysteine, from GO, in a concentration-dependent manner. Inhibition of autolysis of papain by MGX, along with cross-link formation, was detected by SDS/PAGE. Treatment of papain and catS with the dialdehyde o-phthalaldehyde resulted in enzyme inactivation and an intra-molecular active site cysteine-lysine cross-link. These results demonstrate that reactive aldehydes inhibit cysteine proteases by modification of the active site cysteine residue. This process may contribute to the accumulation of modified proteins in tissues of people with diabetes and age-related pathologies, including atherosclerosis, cataract and Alzheimer's disease.

AB - Hyperglycaemia, triose phosphate decomposition and oxidation reactions generate reactive aldehydes in vivo. These compounds react non-enzymatically with protein side chains and N-terminal amino groups to give adducts and cross-links, and hence modified proteins. Previous studies have shown that free or protein-bound carbonyls inactivate glyceraldehyde-3-phosphate dehydrogenase with concomitant loss of thiol groups [Morgan, Dean and Davies (2002) Arch. Biochem. Biophys. 403, 259-269]. It was therefore hypothesized that modification of lysosomal cysteine proteases (and the structurally related enzyme papain) by free and protein-bound carbonyls may modulate the activity of these components of the cellular proteolytic machinery responsible for the removal of modified proteins and thereby contribute to a decreased removal of modified proteins from cells. It is shown that MGX (methylglyoxal), GO (glyoxal) and glycolaldehyde, but not hydroxyacetone and glucose, inhibit catB (cathepsin B), catL (cathepsin L) and catS (cathepsin S) activity in macrophage cell lysates, in a concentration-dependent manner. Protein-bound carbonyls produced similar inhibition with both cell lysates and intact macrophage cells. Inhibition was also observed with papain, with this paralleled by loss of the active site cysteine residue and formation of the adduct species S-carboxymethylcysteine, from GO, in a concentration-dependent manner. Inhibition of autolysis of papain by MGX, along with cross-link formation, was detected by SDS/PAGE. Treatment of papain and catS with the dialdehyde o-phthalaldehyde resulted in enzyme inactivation and an intra-molecular active site cysteine-lysine cross-link. These results demonstrate that reactive aldehydes inhibit cysteine proteases by modification of the active site cysteine residue. This process may contribute to the accumulation of modified proteins in tissues of people with diabetes and age-related pathologies, including atherosclerosis, cataract and Alzheimer's disease.

KW - Animals

KW - Binding Sites

KW - Carbocysteine

KW - Cathepsins

KW - Cattle

KW - Cell Line

KW - Cysteine Endopeptidases

KW - Enzyme Activation

KW - Glycosylation

KW - Glyoxal

KW - Humans

KW - Mice

KW - Molecular Weight

KW - Papain

KW - Protein Binding

KW - Sulfhydryl Compounds

KW - o-Phthalaldehyde

U2 - 10.1042/BJ20060019

DO - 10.1042/BJ20060019

M3 - Journal article

C2 - 16671891

VL - 398

SP - 197

EP - 206

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -

ID: 129671551