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Substrate Recognition by Novel Family of Amino Acid:[Carrier Protein] Ligases

Močibob, Marko; Ivić, Nives; Šubašić, Deni; Luić, Marija; Weygand-Đurašević, Ivana (2011) Substrate Recognition by Novel Family of Amino Acid:[Carrier Protein] Ligases. Croatica Chemica Acta, 84 (2). pp. 149-157. ISSN 0011-1643

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Amino acid:[carrier protein] ligases (aa:CP ligases) are newly discovered homologs of aminoacyl-tRNA synthetases (aaRS) which aminoacylate carrier proteins instead of tRNA. Activated amino acids are attached to prosthetic group of carrier proteins by thioester bond. Weak thioacylation activity was observed for many conventional aaRS. Therefore, different thiols were investigated as substrate analogs for aa:CP ligases. Here we show that coenzyme A, dithiothreitol and cysteine are efficiently aminoacylated by aa:CP ligases. The crystal structure of aa:CP ligase from Bradyrhizobium japonicum in complex with coenzyme A was solved, and revealed that CoA, besides acting as the substrate analog of the carrier protein, also competes with ATP for binding to the active site. aa:CP ligases do not aminoacylate tRNA, although they remarkably resemble catalytic core of atypical seryl-tRNA synthetases (aSerRS) from methanogenic archaea. Since aa:CP ligases lack tRNA-binding domain, fusion proteins of aa:CP ligases and aSerRS tRNA-binding domain were prepared in attempt to restore tRNA aminoacylation activity. Although fusion proteins were able to bind tRNA through appended domain, tRNA could not substitute carrier proteins in aminoacylation reaction.

Item Type: Article
Uncontrolled Keywords: aminoacyl-tRNA synthetase; amino acid:[carrier protein] ligase; carrier protein; coenzyme A; seryl-tRNA synthetase; thioester; tRNA; transfer-rna-synthetases; seryl-transfer-rna; elongation-factor-p; thermus-thermophilus hb8; methanosarcina-barkeri; peptide-synthesis; crystal-structure; coenzyme-a; aminoacylation; model
Subjects: NATURAL SCIENCES > Chemistry
Divisions: Division of Molecular Medicine
Division of Physical Chemistry
Project titleProject leaderProject codeProject type
Strukturna raznolikost seril-tRNA sintetaza i točnost biosinteze proteina[57696] Ivana Weygand Đurašević119-0982913-1358MZOS
Protein-ligand međudjelovanja na atomnoj razini[77430] Marija Luić098-1191344-2943MZOS
Depositing User: Nives Ivić
Date Deposited: 15 Oct 2013 11:58
DOI: 10.5562/cca1818

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