1amu

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PHENYLALANINE ACTIVATING DOMAIN OF GRAMICIDIN SYNTHETASE 1 IN A COMPLEX WITH AMP AND PHENYLALANINE

Authors	Conti, E. Stachelhaus, T. Marahiel, M.A. Brick, P.
Citation	Structural basis for the activation of phenylalanine in the non-ribosomal biosynthesis of gramicidin S.
Release date	1998-07-01
Exp. Method	X-RAY DIFFRACTION
Resolution	1.9 Å
Classification	PEPTIDE SYNTHETASE

Sequence

Chain(s) A B (563 residues): Blast Uniprot EC 5.1.1.11

MVNSSKSILIHAQNKNGTHEEEQYLFAVNNTKAEYPRDKTIHQLFEEQVSKRPNNVAIVCENEQLTYHELNVKANQLARI
FIEKGIGKDTLVGIMMEKSIDLFIGILAVLKAGGAYVPIDIEYPKERIQYILDDSQARMLLTQKHLVHLIHNIQFNGQVE
IFEEDTIKIREGTNLHVPSKSTDLAYVIYTSGTTGNPKGTMLEHKGISNLKVFFENSLNVTEKDRIGQFASISFDASVWE
MFMALLTGASLYIILKDTINDFVKFEQYINQKEITVITLPPTYVVHLDPERILSIQTLITAGSATSPSLVNKWKEKVTYI
NAYGPTETTICATTWVATKETIGHSVPIGAPIQNTQIYIVDENLQLKSVGEAGELCIGGEGLARGYWKRPELTSQKFVDN
PFVPGEKLYKTGDQARWLSDGNIEYLGRIDNQVKIRGHRVELEEVESILLKHMYISETAVSVHKDHQEQPYLCAYFVSEK
HIPLEQLRQFSSEELPTYMIPSYFIQLDKMPLTSNGKIDRKQLPEPDLTFGMRVDYEAPRNEIEETLVTIWQDVLGSHHH
HHH

User comments

Engineered variants

The Bruce Donald lab at Duke University has used computational protein design to change the substrate specificity of this protein from Phe to Leu. They also designed mutants to create activity for 4 charged amino acid substrates; the WT has virtually no detectable (ND) activity for these charged substrates.

Reference:

Cheng-Yu Chen, Ivelin Georgiev, Amy C Anderson, Bruce R Donald
Computational structure-based redesign of enzyme activity.
Proc. Natl. Acad. Sci. U.S.A.: 2009, 106(10);3764-9
[PubMed:19228942] [DOI] (I p)

-- Keedyase 15:23, 14 April 2008 (CET)