Molecular basis for the broad substrate selectivity of a peptide prenyltransferase
| Autor: | Thomas E. Cheatham, Elizabeth Pierce, Vinayak Agarwal, Satish K. Nair, John A. McIntosh, Yue Hao, Daniel R. Roe, Maho Morita, Eric W. Schmidt |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Stereochemistry Amino Acid Motifs Prenyltransferase Peptide Biology Crystallography X-Ray 010402 general chemistry 01 natural sciences Substrate Specificity Peptide substrate Structure-Activity Relationship 03 medical and health sciences Prenylation Catalytic Domain Amino Acid Sequence chemistry.chemical_classification Multidisciplinary fungi food and beverages Active site Regioselectivity Biological Sciences Dimethylallyltranstransferase 0104 chemical sciences 030104 developmental biology Enzyme chemistry biology.protein Peptides Selectivity Protein Binding |
| Zdroj: | Proceedings of the National Academy of Sciences. 113:14037-14042 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1609869113 |
| Popis: | The cyanobactin prenyltransferases catalyze a series of known or unprecedented reactions on millions of different substrates, with no easily observable recognition motif and exquisite regioselectivity. Here we define the basis of broad substrate tolerance for the otherwise uncharacterized TruF family. We determined the structures of the Tyr-prenylating enzyme PagF, in complex with an isoprenoid donor analog and a panel of linear and macrocyclic peptide substrates. Unexpectedly, the structures reveal a truncated barrel fold, wherein binding of large peptide substrates is necessary to complete a solvent-exposed hydrophobic pocket to form the catalytically competent active site. Kinetic, mutational, chemical, and computational analyses revealed the structural basis of selectivity, showing a small motif within peptide substrates that is sufficient for recognition by the enzyme. Attaching this 2-residue motif to two random peptides results in their isoprenylation by PagF, demonstrating utility as a general biocatalytic platform for modifications on any peptide substrate. |
| Databáze: | OpenAIRE |
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