1H, 13C and 15N assignments of the four N-terminal domains of human fibrillin-1
Structure of the Fibrillin-1 N-Terminal Domains Suggests that Heparan Sulfate Regulates the Early Stages of Microfibril Assembly.
FUN-EGF3 region of fibrillin-1, UNP residues 45-178
SARGGGGHDALKGPNVCGSRYNAYCCPGWKTLPGGNQCIVPICRHSCGDGFCSRPNMCTCPSGQIAPSCGSRSIQHCNIR CMNGGSCSDDHCLCQKGYIGTHCGQPVCESGCLNGGRCVAPNRCACTYGFTGPQCE
Cells were also transformed with the pREP4 plasmid, enabling inducible expression
Chemical shift assignments for the FUN-EGF3 recombinant fragment of human fibrillin-1
An ensemble of 100 structures was generated. Floating chirality of prochiral groups was used for stereospecific assignment., An initial ensemble of 200 structures was using simulated annealing from an extended template using NOE-derived distance restraints with stereospecific assignments. Coordinates of the 10 lowest-energy structures were then used as templates for refinement calculations, including the 'Rama' and 'HBDB' database potentials. The resulting 10 lowest-energy structures were then refined further using RDCs. Separate alignment tensors were used for the two halves of the molecule ('ne1' and 'e2e3'). Alignment tensor parameters were estimated using partially refined structures. The 20 lowest-structures from the final ensemble of 200 structures were selected as the representative structure ensemble.
simulated annealing, torsion angle dynamics
Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
Schwieters, Kuszewski, Tjandra and Clore
home-built home-built using GE Omega software
RGGGGHDALKGPNVCGSRYNAYCCPGWKTLPGGNQCIVPICRHSCGDGFCSRPNMCTCPSGQIAPSCGSRSIQHCNIRCM NGGSCSDDHCLCQKGYIGTHCGQPVCESGCLNGGRCVAPNRCACTYGFTGPQCE