Simultaneous recognition of HIV-1 TAR RNA bulge and loop sequences by cyclic peptide mimic of Tat protein
Simultaneous recognition of HIV-1 TAR RNA bulge and loop sequences by cyclic peptide mimics of Tat protein
BIV TAR and cyclic peptide mimicking BIV Tat
THE L-22 PEPTIDE WAS SYNTHESIZED BY SOLID PHASE SYNTHESIS AND PURIFIED BY REVERSE PHASE HPLC
RNA WAS TRANSCRIBED IN VITRO FROM DNA OLIGONUCLEOTIDE TEMPLATES WITH T7 RNA POLYMERASE PURIFIED IN HOUSE WITH UNLABELED OR 13C/15N ENRICHED NTPS (ISOTEC) AND PURIFIED BY DENATURING PAGE.
Lowest energy, least restraint violations
3D-13C HMQC NOESY, 3D HCCH TOCSY, IPAP-HSQC
F1fF2f-type noesy, F1fF2f-type tocsy, 2D HCCH-COSY
Structures of the HIV-1 TAR RNA/L-22 complex were calculated with Xplor-NIH. Backbone dihedral angle restraints for the peptide were estimated using chemical shift data and TALOS. Structures were originally calculated without RDCs to test for convergence and adherence to the NOE data. RDC restraints were then applied as susceptibility anisotropy restraints with a harmonic potential well.
1.0mM HIV-1 TAR RNA-1, 1.0mM L-22 CYCLIC PEPTIDE-2, 100% D2O
1.0mM HIV-1 TAR RNA-3, 1.0mM L-22 CYCLIC PEPTIDE-4, 90 % H2O, 10% D2O
1.0mM [U-2H] HIV-1 TAR RNA-9, 1.0mM L-22 CYCLIC PEPTIDE-10, 100% D2O
C.D. Schwieters, J.J. Kuszewski, N. Tjandra and G.M. Clore
Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
PEPTIDOMIMETICS, PEPTIDE STRUCTURE, RNA RECOGNITION, IMMUNODEFICIENCY VIRUS, TAR RNA, mimic of RNA binding protein, RNA binding protein-RNA COMPLEX