title: Rational protein engineering in action: the first crystal structure of a phenylalanine tRNA synthetase from Staphylococcus haemolyticus.
abstract: In this article, we describe for the first time the high-resolution crystal structure of a phenylalanine tRNA synthetase from the pathogenic bacterium Staphylococcus haemolyticus. We demonstrate the subtle yet important structural differences between this enzyme and the previously described Thermus thermophilus ortholog. We also explain the structure-activity relationship of several recently reported inhibitors. The native enzyme crystals were of poor quality--they only diffracted X-rays to 3-5A resolution. Therefore, we have executed a rational surface mutagenesis strategy that has yielded crystals of this 2300-amino acid multidomain protein, diffracting to 2A or better. This methodology is discussed and contrasted with the more traditional domain truncation approach.
authors: Pfizer Global Research & Development, Michigan Laboratories, Ann Arbor, 2800 Plymouth Road, Ann Arbor, MI 48105, USA. artem@xtals.org
authors: Artem G
authors: AG
authors: Evdokimov
authors: Marlene
authors: M
authors: Mekel
authors: Kim
authors: K
authors: Hutchings
authors: Lakshmi
authors: L
authors: Narasimhan
authors: Tod
authors: T
authors: Holler
authors: Teresa
authors: T
authors: McGrath
authors: Bryan
authors: B
authors: Beattie
authors: Eric
authors: E
authors: Fauman
authors: Chunhong
authors: C
authors: Yan
authors: Holly
authors: H
authors: Heaslet
authors: Richard
authors: R
authors: Walter
authors: Barry
authors: B
authors: Finzel
authors: Jeffrey
authors: J
authors: Ohren
authors: Patrick
authors: P
authors: McConnell
authors: Timothy
authors: T
authors: Braden
authors: Fang
authors: F
authors: Sun
authors: Cindy
authors: C
authors: Spessard
authors: Craig
authors: C
authors: Banotai
authors: Loola
authors: L
authors: Al-Kassim
authors: Weijun
authors: W
authors: Ma
authors: Paul
authors: P
authors: Wengender
authors: Denis
authors: D
authors: Kole
authors: Norman
authors: N
authors: Garceau
authors: Peter
authors: P
authors: Toogood
authors: Jia
authors: J
authors: Liu
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018360:Crystallography, X-Ray / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016296:Mutagenesis; D010652:Phenylalanine-tRNA Ligase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D041162:Staphylococcus haemolyticus / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D010652:Phenylalanine-tRNA Ligase