4KOT

Crystal structure of a GNAT superfamily acetyltransferase PA4794 in complex with Cefotaxime

Summary for 4KOT

• Entry DOI: 10.2210/pdb4kot/pdb
• Related: 3PGP 4KLV 4KLW 4KOR 4KOS 4KOU 4KOV 4KOW 4KOX 4KOY 4KUA 4KUB
• Descriptor: Uncharacterized protein, SULFATE ION, (6R,7R)-3-(acetyloxymethyl)-7-[[(2Z)-2-(2-amino-1,3-thiazol-4-yl)-2-methoxyimino-ethanoyl]amino]-8-oxo-5-thia-1-azabicy clo[4.2.0]oct-2-ene-2-carboxylic acid, ... (5 entities in total)
• Functional Keywords: structural genomics, psi-biology, midwest center for structural genomics, mcsg, transferase
• Biological source: Pseudomonas aeruginosa
• Total number of polymer chains: 1
• Total formula weight: 19150.66

Authors

Majorek, K.A.,Chruszcz, M.,Joachimiak, A.,Minor, W.,Midwest Center for Structural Genomics (MCSG) (deposition date: 2013-05-12, release date: 2013-06-05, Last modification date: 2023-09-20)

Primary citation

Majorek, K.A.,Kuhn, M.L.,Chruszcz, M.,Anderson, W.F.,Minor, W.
Structural, Functional, and Inhibition Studies of a Gcn5-related N-Acetyltransferase (GNAT) Superfamily Protein PA4794: A NEW C-TERMINAL LYSINE PROTEIN ACETYLTRANSFERASE FROM PSEUDOMONAS AERUGINOSA.
J.Biol.Chem., 288:30223-30235, 2013

PubMed Abstract: The Gcn5-related N-acetyltransferase (GNAT) superfamily is a large group of evolutionarily related acetyltransferases, with multiple paralogs in organisms from all kingdoms of life. The functionally characterized GNATs have been shown to catalyze the transfer of an acetyl group from acetyl-coenzyme A (Ac-CoA) to the amine of a wide range of substrates, including small molecules and proteins. GNATs are prevalent and implicated in a myriad of aspects of eukaryotic and prokaryotic physiology, but functions of many GNATs remain unknown. In this work, we used a multi-pronged approach of x-ray crystallography and biochemical characterization to elucidate the sequence-structure-function relationship of the GNAT superfamily member PA4794 from Pseudomonas aeruginosa. We determined that PA4794 acetylates the Nε amine of a C-terminal lysine residue of a peptide, suggesting it is a protein acetyltransferase specific for a C-terminal lysine of a substrate protein or proteins. Furthermore, we identified a number of molecules, including cephalosporin antibiotics, which are inhibitors of PA4794 and bind in its substrate-binding site. Often, these molecules mimic the conformation of the acetylated peptide product. We have determined structures of PA4794 in the apo-form, in complexes with Ac-CoA, CoA, several antibiotics and other small molecules, and a ternary complex with the products of the reaction: CoA and acetylated peptide. Also, we analyzed PA4794 mutants to identify residues important for substrate binding and catalysis.

PubMed: 24003232
DOI: 10.1074/jbc.M113.501353

Experimental method

X-RAY DIFFRACTION (1.55 Å)