Adaptation of an L-Proline Adenylation Domain to Use 4-Propyl-L-Proline in the Evolution of Lincosamide Biosynthesis

0425633 - MBÚ 2014 RIV US eng J - Journal Article
Kadlčík, Stanislav - Kučera, Tomáš - Chalupská, Dominika - Gažák, Radek - Koběrská, Markéta - Ulanová, Dana - Kopecký, Jan - Kutejová, Eva - Najmanová, Lucie - Janata, Jiří
Adaptation of an L-Proline Adenylation Domain to Use 4-Propyl-L-Proline in the Evolution of Lincosamide Biosynthesis.
PLoS ONE. Roč. 8, č. 12 (2013). ISSN 1932-6203. E-ISSN 1932-6203
R&D Projects: GA MŠMT(CZ) EE2.3.20.0055; GA MŠMT(CZ) EE2.3.30.0003; GA MŠMT ED1.1.00/02.0109
Institutional support: RVO:61388971
Keywords : NONRIBOSOMAL PEPTIDE SYNTHETASES * GENE-CLUSTER * BIOCHEMICAL-CHARACTERIZATION
Subject RIV: EE - Microbiology, Virology
Impact factor: 3.534, year: 2013

Clinically used lincosamide antibiotic lincomycin incorporates in its structure 4-propyl-L-proline (PPL), an unusual amino acid, while celesticetin, a less efficient related compound, makes use of proteinogenic L-proline. Biochemical characterization, as well as phylogenetic analysis and homology modelling combined with the molecular dynamics simulation were employed for complex comparative analysis of the orthologous protein pair LmbC and CcbC from the biosynthesis of lincomycin and celesticetin, respectively. The analysis proved the compared proteins to be the standalone adenylation domains strictly preferring their own natural substrate, PPL or L-proline. The LmbC substrate binding pocket is adapted to accomodate a rare PPL precursor. When compared with L-proline specific ones, several large amino acid residues were replaced by smaller ones opening a channel which allowed the alkyl side chain of PPL to be accommodated. One of the most important differences, that of the residue corresponding to V306 in CcbC changing to G308 in LmbC, was investigated in vitro and in silico.
Permanent Link: http://hdl.handle.net/11104/0231486