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NMR Solution Structures of 3-oxo-butyl-ACP, an intermediate mimic from the actinorhodin polyketide synthase in Streptomyces coelicolor
Authors
Crump, M.P., Evans, S.E., Williams, C.
Assembly
3-oxo-butyl-ACP
Entity
1. 3-oxo-butyl-ACP (polymer, Thiol state: not present), 86 monomers, 9232.119 Da Detail

MATLLTTDDL RRALVESAGE TDGTDLSGDF LDLRFEDIGY DSLALMETAA RLESRYGVSI PDDVAGRVDT PRELLDLING ALAEAA


2. N~3~-[(2S)-2-hydroxy-3,3-dimethyl-4-(phosphonooxy)butanoyl]-N-{2-[(3-oxobutyl)sulfanyl]ethyl}-beta-alaninamide (non-polymer), 428.438 Da
Total weight
9660.557 Da
Max. entity weight
9232.119 Da
Source organism
Streptomyces coelicolor
Exptl. method
solution NMR
Refine. method
simulated annealing
Data set
assigned_chemical_shifts
Chem. Shift Complete
Sequence coverage: 98.8 %, Completeness: 97.4 %, Completeness (bb): 98.0 % Detail

Polymer type: polypeptide(L)

Total1H13C15N
All97.4 % (887 of 911)96.8 % (447 of 462)97.8 % (356 of 364)98.8 % (84 of 85)
Backbone98.0 % (502 of 512)97.7 % (173 of 177)98.0 % (246 of 251)98.8 % (83 of 84)
Sidechain96.9 % (463 of 478)96.1 % (274 of 285)97.9 % (188 of 192)100.0 % (1 of 1)
Aromatic100.0 % (36 of 36)100.0 % (18 of 18)100.0 % (18 of 18)
Methyl98.3 % (118 of 120)96.7 % (58 of 60)100.0 % (60 of 60)

1. 3-oxo-butyl-ACP

MATLLTTDDL RRALVESAGE TDGTDLSGDF LDLRFEDIGY DSLALMETAA RLESRYGVSI PDDVAGRVDT PRELLDLING ALAEAA

Sample

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5 (±0.2), Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 3-oxo-butyl group and 4'PP chain were not isotopically labelled.


#NameIsotope labelingTypeConcentration
1ACPnatural abundance1.0 ~ 2.0 (±0.2) mM
2potassium phosphatenatural abundance20 (±2.0) mM
3sodium azidenatural abundance1 (±0.1) mM
4H2Onatural abundance95 %
5D2Onatural abundance5 %

LACS Plot; CA
Referencing offset: -0.29 ppm, Outliers: 1 Detail
LACS Plot; CB
Referencing offset: -0.29 ppm, Outliers: 1 Detail
LACS Plot; HA
Referencing offset: -0.12 ppm, Outliers: 1 Detail
LACS Plot; CO
Referencing offset: -0.04 ppm, Outliers: 3 Detail
Protein Blocks Logo
Calculated from 20 models in PDB: 2KGD, Strand ID: A Detail


Release date
2009-04-21
Citation
Probing the Interactions of early polyketide intermediates with the Actinorhodin ACP from S. coelicolor A3(2)
Evans, S.E., Williams, C., Arthur, C.J., Ploskon, E., Wattana-Amorn, P., Cox, R.J., Crosby, J., Willis, C.L., Simpson, T.J., Crump, M.P.
J. Mol. Biol. (2009), 389, 511-528, PubMed 19361520 , DOI 10.1016/j.jmb.2009.03.072 ,
Related entities 1. 3-oxo-butyl-ACP, : 1 : 10 : 3 : 64 entities Detail
Experiments performed 12 experiments Detail
NMR combined restraints 4 contents Detail
Keywords ACP, Actinorhodin, Coelicolor, diketide, polyketide mimic