title: Michael addition of dehydroalanine-containing MAPK peptides to catalytic lysine inhibits the activity of phosphothreonine lyase.
abstract: The phosphothreonine lyases OspF and SpvC irreversibly inactivate host dual-phosphorylated mitogen-activated protein kinases (MAPKs) [pThr-X-pTyr motif] through β-elimination. We found that dual-phosphorylated (pSer-X-pTyr) MAPK substrate peptides and their resulting catalytic products cross-link to OspF and SpvC. Mass spectrometry results revealed that these linkages form between lysine, which acts as a general base, and dehydroalanine (Dha) on catalytic products. The nucleophilic addition efficiency is dependent on the K136 residue being in a deprotonated state. Peptide cross-linking inhibits the activity of SpvC and blocks the inactivation of MAPK signaling by SpvC. Small compounds mimicking these sequences may act as phosphothreonine lyase inhibitors.
authors.affiliation: The State Key Laboratory Breeding Base of Bioresources and Eco-environments, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Molecular Developmental Biology, School of Life Sciences, Southwest University, Beibei, 400715 Chongqing, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: The State Key Laboratory Breeding Base of Bioresources and Eco-environments, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Molecular Developmental Biology, School of Life Sciences, Southwest University, Beibei, 400715 Chongqing, China.
authors.forename: Ru
authors.initials: R
authors.lastname: Yang
authors.affiliation: The State Key Laboratory Breeding Base of Bioresources and Eco-environments, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Molecular Developmental Biology, School of Life Sciences, Southwest University, Beibei, 400715 Chongqing, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Huang
authors.affiliation: The State Key Laboratory Breeding Base of Bioresources and Eco-environments, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Molecular Developmental Biology, School of Life Sciences, Southwest University, Beibei, 400715 Chongqing, China.
authors.forename: Qiujin
authors.initials: Q
authors.lastname: Liang
authors.affiliation: The State Key Laboratory Breeding Base of Bioresources and Eco-environments, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Molecular Developmental Biology, School of Life Sciences, Southwest University, Beibei, 400715 Chongqing, China.
authors.forename: Yanmin
authors.initials: Y
authors.lastname: Guo
authors.affiliation: The State Key Laboratory Breeding Base of Bioresources and Eco-environments, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Molecular Developmental Biology, School of Life Sciences, Southwest University, Beibei, 400715 Chongqing, China.
authors.forename: Weixiang
authors.initials: W
authors.lastname: Bian
authors.affiliation: The State Key Laboratory Breeding Base of Bioresources and Eco-environments, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Molecular Developmental Biology, School of Life Sciences, Southwest University, Beibei, 400715 Chongqing, China.
authors.forename: Lingfei
authors.initials: L
authors.lastname: Luo
authors.affiliation: The State Key Laboratory Breeding Base of Bioresources and Eco-environments, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Molecular Developmental Biology, School of Life Sciences, Southwest University, Beibei, 400715 Chongqing, China. Electronic address: htli@swu.edu.cn.
authors.forename: Hongtao
authors.initials: H
authors.lastname: Li
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives* / Q000737:chemistry; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D055162:Biocatalysis; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D008190:Lyases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry* / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000737:chemistry* / Q000378:metabolism; D057786:Peptidomimetics / Q000737:chemistry / Q000494:pharmacology*; D010766:Phosphorylation; D010769:Phosphothreonine / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D057786:Peptidomimetics; D010769:Phosphothreonine; C015102:dehydroalanine; D020928:Mitogen-Activated Protein Kinases; D008190:Lyases; D008239:Lysine; D000409:Alanine
keywords: Cross-linking; Dehydroalanine; Effector; Inhibit; Mitogen-activated protein kinase; Phosphothreonine lyase