H-1-NMR STUDY OF THE SOLUTION STRUCTURE OF AC-AMP2, A SUGAR BINDING ANTIMICROBIAL PROTEIN ISOLATED FROM AMARANTHUS-CAUDATUS

Citation
Jc. Martins et al., H-1-NMR STUDY OF THE SOLUTION STRUCTURE OF AC-AMP2, A SUGAR BINDING ANTIMICROBIAL PROTEIN ISOLATED FROM AMARANTHUS-CAUDATUS, Journal of Molecular Biology, 258(2), 1996, pp. 322-333
Citations number
39
Categorie Soggetti
Biology
ISSN journal
00222836
Volume
258
Issue
2
Year of publication
1996
Pages
322 - 333
Database
ISI
SICI code
0022-2836(1996)258:2<322:HSOTSS>2.0.ZU;2-C
Abstract
The conformation in water of antimicrobial protein 2 from Amaranthus c audatus (Ac-AMP2) was determined using H-1 NMR, DIANA and restrained m olecular modeling. Ac-AMP2 is a 30 amino acid residue, lectin-like pro tein that specifically binds to chitin, a polymer of beta-1,4-N-acetyl -D-glucosamine. After sequence specific resonance assignments, a total of 198 distance restraints were collected from 2D NOESY buildup spect ra at 500 MHz at pH 2, supplemented by a 2D NOESY spectrum at 600 MHz. The location of the three previously unassigned disulfide bridges was determined from preliminary DIANA structures, using a statistical ana lysis of intercystinyl distances. The solution structure of Ac-AMP2 is presented as a set of 26 DIANA structures, further refined by restrai ned molecular dynamics using a simulated annealing protocol in the AMB ER force field, with a backbone r.m.s.d. for the well defined Glu3-Cys 28 segment of 0.69(+/-0.12) Angstrom. The main structural element is a n antiparallel beta-sheet from Met13 to Lys23 including a beta(I)-turn over Gln17-Phe18 with a beta bulge at Gly19. Ln addition, a beta'(I) turn over Arg6-Gly7, a beta'(III) turn over Ser11-Gly12 and a helical turn from Gly24 to Cys28 are identified. This structure is very simila r to the equivalent regions of the X-ray structure of wheat germ agglu tinin and the NMR structure of hevein. (C) 1996 Academic Press Limited