Teicoplanin Nonribosomal Peptide Synthetase Is Unable to Incorporate Alpha-Ketoacid Building Blocks DOI

Minuri S. Ratnayake,

Xinyun Jian, Julien Tailhades

и другие.

Biochemistry, Год журнала: 2025, Номер unknown

Опубликована: Апрель 11, 2025

Glycopeptide antibiotics (GPAs) are a vital class of nonribosomal peptides used as therapies last resort to treat infections by multidrug-resistant bacteria. These peptide assembled synthetases (NRPSs), modular megasynthases central the biosynthesis wide range natural products. The adenylation (A) domains NRPSs involved in selection and activation amino acid building blocks forming these products, with their subsequent loading onto neighboring carrier protein for incorporation into growing chain. This makes A-domains gatekeepers specificity biosynthesis, further studies needed reveal how this is enforced at all stages catalysis. first block found GPAs diverse can comprise an acid, ketoacid, or mixtures both, which suggests that responsible selecting residues also incorporate non-amino substrates. In study, we explored acceptance such substrates initiation module teicoplanin NRPS. Our vitro assays demonstrated A-domain possesses unexpected preference activating ketoacids over native substrate l-Hpg. However, only (d/l)-Hpg related acids were able be loaded domain during thioesterification step. We characterized structure from complex d-4-hydroxyphenylglycine (d-Hpg), revealed alterations positioning carboxylate help explain high levels pyrophosphate release seen acid. combination extensive molecular dynamics simulations, data suggest ketoacid GPA likely performed after NRPS highlight importance considering both reactions when investigating selectivity biosynthesis.

Язык: Английский

Activity Regulation and Conformation Response of Janus Kinase 3 Mediated by Phosphorylation: Exploration from Correlation Network Analysis and Markov Model DOI

Jian‐Zhong Chen,

Jian Wang, Wanchun Yang

и другие.

Journal of Chemical Information and Modeling, Год журнала: 2025, Номер unknown

Опубликована: Апрель 8, 2025

The activity of the enzyme JAK3 is modulated by tyrosine phosphorylation, yet underlying molecular details remain not fully understood. In this study, we employed a GaMD trajectory-based Markov model and correlation network analysis (CNA) to investigate impact single phosphorylation (SP) at Y980 (pY980) double (DP) Y980/Y981 (pY980/pY981) on conformational dynamics bound inhibitors IZA MI1. indicated that both SP DP result in fewer states significantly influence P-loop, αC-helix, loop1-loop3, while maintaining hinge region's high rigidity. CNA findings revealed alters communication among different structural regions JAK3, providing rational explanation for how affects distant P-loop loop1-loop3. Moreover, changes mediated further affect interactions between hot spots (L828, V836, E903, Y904, L905, L956) JAK3. This work offers valuable theoretical insights into mechanisms regulate activity.

Язык: Английский

Процитировано

0
Teicoplanin Nonribosomal Peptide Synthetase Is Unable to Incorporate Alpha-Ketoacid Building Blocks DOI

Minuri S. Ratnayake,

Xinyun Jian, Julien Tailhades

и другие.

Biochemistry, Год журнала: 2025, Номер unknown

Опубликована: Апрель 11, 2025

Glycopeptide antibiotics (GPAs) are a vital class of nonribosomal peptides used as therapies last resort to treat infections by multidrug-resistant bacteria. These peptide assembled synthetases (NRPSs), modular megasynthases central the biosynthesis wide range natural products. The adenylation (A) domains NRPSs involved in selection and activation amino acid building blocks forming these products, with their subsequent loading onto neighboring carrier protein for incorporation into growing chain. This makes A-domains gatekeepers specificity biosynthesis, further studies needed reveal how this is enforced at all stages catalysis. first block found GPAs diverse can comprise an acid, ketoacid, or mixtures both, which suggests that responsible selecting residues also incorporate non-amino substrates. In study, we explored acceptance such substrates initiation module teicoplanin NRPS. Our vitro assays demonstrated A-domain possesses unexpected preference activating ketoacids over native substrate l-Hpg. However, only (d/l)-Hpg related acids were able be loaded domain during thioesterification step. We characterized structure from complex d-4-hydroxyphenylglycine (d-Hpg), revealed alterations positioning carboxylate help explain high levels pyrophosphate release seen acid. combination extensive molecular dynamics simulations, data suggest ketoacid GPA likely performed after NRPS highlight importance considering both reactions when investigating selectivity biosynthesis.

Язык: Английский

Процитировано

0