Alkylation of protein by methyl methanesulfonate and 1-methyl-1-nitrosourea in vitro

Paik, Woon Ki; DiMaria, Peter; Kim, Sangduk; Magee, Peter; Lotlikar, Prabhakar D.

doi:10.1016/0304-3835(84)90056-9

Cited by 10 publications

(6 citation statements)

References 20 publications

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“…In contrast to UV, we did not obtain evidence for direct modification of SH groups of tyrosine phosphatases by MMS (40a). In fact, in vitro MMS preferentially modifies lysine and histidine residues, rather than cysteines (60). Nevertheless, we found the redox state of the cell very critical for induction of the MMS response.…”

Section: Discussionmentioning

confidence: 60%

The Level of Intracellular Glutathione Is a Key Regulator for the Induction of Stress-Activated Signal Transduction Pathways Including Jun N-Terminal Protein Kinases and p38 Kinase by Alkylating Agents

Wilhelm

Bender²,

Knebel³

et al. 1997

Molecular and Cellular Biology

216

152

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Section: Discussionmentioning

confidence: 60%

The Level of Intracellular Glutathione Is a Key Regulator for the Induction of Stress-Activated Signal Transduction Pathways Including Jun N-Terminal Protein Kinases and p38 Kinase by Alkylating Agents

Wilhelm

Bender²,

Knebel³

et al. 1997

Molecular and Cellular Biology

216

152

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“…It is also important to note that solely depleting GSH (with BSO or NRF2 knockdown) was not sufficient to affect cell survival (up to 72 h) or to cause ER stress; the presence of alkylating agent to trigger damage at the same time was required. MMS has been described as being able to directly alkylate proteins (47, 48) as well as GSH (49). In addition, alkylation of ER proteins such as glucose-regulated protein 94 (GRP94), calreticulin, valosin containing protein (VCP) and HSP90 has also been reported in CDDP-treated Cochlear and kidney cells (50).…”

Section: Discussionmentioning

confidence: 99%

Alkylating Agent–Induced NRF2 Blocks Endoplasmic Reticulum Stress–Mediated Apoptosis via Control of Glutathione Pools and Protein Thiol Homeostasis

Zanotto-Filho

Masamsetti

Loranc³

et al. 2016

Molecular Cancer Therapeutics

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Alkylating agents are a commonly used cytotoxic class of anticancer drugs. Understanding the mechanisms whereby cells respond to these drugs is key to identify means to improve therapy while reducing toxicity. By integrating genome-wide gene expression profiling, protein analysis and functional cell validation, we herein demonstrated a direct relationship between NRF2 and Endoplasmic Reticulum (ER) stress pathways in response to alkylating agents, which is coordinated by the availability of glutathione (GSH) pools. GSH is essential for both drug detoxification and protein thiol homeostasis within the ER, thus inhibiting ER stress induction and promoting survival; an effect independent of its antioxidant role. NRF2 accumulation induced by alkylating agents resulted in increased GSH synthesis via GCLC/GCLM enzyme, and interfering with this NRF2 response by either NRF2 knockdown or GCLC/GCLM inhibition with buthionine sulfoximine (BSO) caused accumulation of damaged proteins within the ER, leading to PERK-dependent apoptosis. Conversely, upregulation of NRF2, through KEAP1 depletion or NRF2-myc overexpression, or increasing GSH levels with N-acetylcysteine (NAC) or glutathione-ethyl-ester (GSH-E), decreased ER stress and abrogated alkylating agents-induced cell death. Based on these results, we identified a subset of lung and head-and-neck carcinomas with mutations in either KEAP1 or NRF2/NFE2L2 genes that correlate with NRF2 targets overexpression and poor survival. In KEAP1 mutant cancer cells, NRF2 knockdown and GSH depletion increased cell sensitivity via ER stress induction in a mechanism specific to alkylating drugs. Overall, we show that the NRF2-GSH influence on ER homeostasis implicates defects in NRF2-GSH or ER stress machineries as affecting alkylating therapy toxicity.

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“…However, protein alkylation by MNU and MMS has not been investigated as thoroughly. According to the pioneering work of Paik and colleagues on the reaction of MMS and MNU with yeast cytochrome c specific protein methylase, only methylated protein adducts were found following MMS treatment 10. However, no detailed information was provided on possible protein adducts of MNU.…”

mentioning

confidence: 99%

Differential adduction of proteins vs. deoxynucleosides by methyl methanesulfonate and 1‐methyl‐1‐nitrosourea in vitro

Zhang¹,

Bartels²,

Pottenger³

et al. 2005

Rapid Comm Mass Spectrometry

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The reactions of two model mutagenic and carcinogenic alkylating agents, N-methyl-N-nitrosourea (MNU) and methyl methanesulfonate (MMS), with proteins and deoxynucleosides in vitro, were investigated. The protein work used an approach involving trypsin digestion and high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS). This technique permitted identification of the specific location of protein adduction by both MNU and MMS with commercial apomyoglobin and human hemoglobin, under physiological conditions. MNU treatment resulted in predominantly carbamoylation adducts on the proteins, but in contrast only methylated protein adducts were found following treatment with MMS. Further analyses, using TurboSequest, and the Scoring Algorithm for Spectral Analysis (SALSA), revealed that MNU carbamoylation was specific for modification of either the N-terminal valine or the free amino group in lysine residues of apomyglobin and human hemoglobin. However, MMS methylation modified the N-terminal valine and histidine residues of the proteins. Despite their clear differences in protein modifications, MNU and MMS formed qualitatively the same methylated deoxynucleoside adduct profiles with all four deoxynucleosides in vitro under physiological conditions. In light of their different biological potencies, where MMS is considered a 'super clastogen' while MNU is a 'super mutagen', these differences in reaction products with proteins vs. deoxynucleosides may indicate that these two model alkylating agents work via different mechanisms to produce their mutagenic and carcinogenic effects.

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Alkylation of protein by methyl methanesulfonate and 1-methyl-1-nitrosourea in vitro

Cited by 10 publications

References 20 publications

The Level of Intracellular Glutathione Is a Key Regulator for the Induction of Stress-Activated Signal Transduction Pathways Including Jun N-Terminal Protein Kinases and p38 Kinase by Alkylating Agents

The Level of Intracellular Glutathione Is a Key Regulator for the Induction of Stress-Activated Signal Transduction Pathways Including Jun N-Terminal Protein Kinases and p38 Kinase by Alkylating Agents

Alkylating Agent–Induced NRF2 Blocks Endoplasmic Reticulum Stress–Mediated Apoptosis via Control of Glutathione Pools and Protein Thiol Homeostasis

Differential adduction of proteins vs. deoxynucleosides by methyl methanesulfonate and 1‐methyl‐1‐nitrosourea in vitro

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