7UX8

Crystal structure of MfnG, an L- and D-tyrosine O-methyltransferase from the marformycin biosynthesis pathway of Streptomyces drozdowiczii, with SAH and L-Tyrosine bound at 1.4 A resolution (P212121 - form II)

  • Classification: TRANSFERASE
  • Organism(s): Streptomyces drozdowiczii
  • Expression System: Escherichia coli BL21(DE3)
  • Mutation(s): No 

  • Deposited: 2022-05-05 Released: 2022-09-28 
  • Deposition Author(s): Miller, M.D., Wu, K.-L., Xu, W., Xiao, H., Philips Jr., G.N.
  • Funding Organization(s): National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS), National Institutes of Health/National Cancer Institute (NIH/NCI), National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID), Cancer Prevention and Research Institute of Texas (CPRIT), Robert A. Welch Foundation, Department of Defense (DOD, United States), National Science Foundation (NSF, United States)

Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.40 Å
  • R-Value Free: 0.172 
  • R-Value Work: 0.153 
  • R-Value Observed: 0.154 

wwPDB Validation   3D Report Full Report


Ligand Structure Quality Assessment 


This is version 1.2 of the entry. See complete history


Literature

Expanding the eukaryotic genetic code with a biosynthesized 21st amino acid.

Wu, K.L.Moore, J.A.Miller, M.D.Chen, Y.Lee, C.Xu, W.Peng, Z.Duan, Q.Phillips Jr., G.N.Uribe, R.A.Xiao, H.

(2022) Protein Sci 31: e4443-e4443

  • DOI: https://doi.org/10.1002/pro.4443
  • Primary Citation of Related Structures:  
    7UX6, 7UX7, 7UX8

  • PubMed Abstract: 

    Genetic code expansion technology allows for the use of noncanonical amino acids (ncAAs) to create semisynthetic organisms for both biochemical and biomedical applications. However, exogenous feeding of chemically synthesized ncAAs at high concentrations is required to compensate for the inefficient cellular uptake and incorporation of these components into proteins, especially in the case of eukaryotic cells and multicellular organisms. To generate organisms capable of autonomously biosynthesizing an ncAA and incorporating it into proteins, we have engineered a metabolic pathway for the synthesis of O-methyltyrosine (OMeY). Specifically, we endowed organisms with a marformycins biosynthetic pathway-derived methyltransferase that efficiently converts tyrosine to OMeY in the presence of the co-factor S-adenosylmethionine. The resulting cells can produce and site-specifically incorporate OMeY into proteins at much higher levels than cells exogenously fed OMeY. To understand the structural basis for the substrate selectivity of the transferase, we solved the X-ray crystal structures of the ligand-free and tyrosine-bound enzymes. Most importantly, we have extended this OMeY biosynthetic system to both mammalian cells and the zebrafish model to enhance the utility of genetic code expansion. The creation of autonomous eukaryotes using a 21st amino acid will make genetic code expansion technology more applicable to multicellular organisms, providing valuable vertebrate models for biological and biomedical research.


  • Organizational Affiliation

    Department of Chemistry, Rice University, Houston, Texas, USA.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
MfnG
A, B
384Streptomyces drozdowicziiMutation(s): 0 
UniProt
Find proteins for A0A0D4WTP2 (Streptomyces drozdowiczii)
Explore A0A0D4WTP2 
Go to UniProtKB:  A0A0D4WTP2
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupA0A0D4WTP2
Sequence Annotations
Expand
  • Reference Sequence
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.40 Å
  • R-Value Free: 0.172 
  • R-Value Work: 0.153 
  • R-Value Observed: 0.154 
  • Space Group: P 21 21 21
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 60.837α = 90
b = 87.167β = 90
c = 134.91γ = 90
Software Package:
Software NamePurpose
PHENIXrefinement
XDSdata reduction
XDSdata scaling
PDB_EXTRACTdata extraction
PHENIXphasing

Structure Validation

View Full Validation Report



Ligand Structure Quality Assessment 


Entry History & Funding Information

Deposition Data


Funding OrganizationLocationGrant Number
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)United StatesR01-GM115261
National Institutes of Health/National Cancer Institute (NIH/NCI)United StatesR01-CA217255
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)United StatesR35-GM133706
National Institutes of Health/National Cancer Institute (NIH/NCI)United StatesR21-CA255894
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)United StatesR01-AI165079
Cancer Prevention and Research Institute of Texas (CPRIT)United StatesRR170014
Robert A. Welch FoundationUnited StatesC-1970
Department of Defense (DOD, United States)United StatesW81XWH-21-1-0789
National Science Foundation (NSF, United States)United StatesSTC 1231306

Revision History  (Full details and data files)

  • Version 1.0: 2022-09-28
    Type: Initial release
  • Version 1.1: 2022-10-12
    Changes: Database references
  • Version 1.2: 2023-10-18
    Changes: Data collection, Refinement description