AUTOMATED NOESY INTERPRETATION WITH AMBIGUOUS DISTANCE RESTRAINTS - THE REFINED NMR SOLUTION STRUCTURE OF THE PLECKSTRIN HOMOLOGY DOMAIN FROM BETA-SPECTRIN
M. Nilges et al., AUTOMATED NOESY INTERPRETATION WITH AMBIGUOUS DISTANCE RESTRAINTS - THE REFINED NMR SOLUTION STRUCTURE OF THE PLECKSTRIN HOMOLOGY DOMAIN FROM BETA-SPECTRIN, Journal of Molecular Biology, 269(3), 1997, pp. 408-422
We have used a novel, largely automated, calculation method to refine
the NMR solution structure of the pleckstrin homology domain of beta-s
pectrin. The method is called ARIA for Ambiguous Restraints for Iterat
ive Assignment. The starting Feint for ARIA is an almost complete assi
gnment of the proton chemical shifts, and a list of partially assigned
NOEs, mostly sequential and secondary structure NOEs. The restraint l
ist is then augmented by automatically interpreting peak lists generat
ed by automated peak-picking. The central task of ARIA is the assignme
nt of ambiguous NOEs during the structure calculation using a combinat
ion of ambiguous distance restraints and an iterative assignment strat
egy. Ln addition, ARIA calibrates ambiguous NOEs to derive distance re
straints, merges overlapping data sets to remove duplicate information
, and uses empirical rules to identify erroneous peaks. While the dist
ance restraints for the structure calculations were exclusively extrac
ted from homonuclear 2D experiments, ARIA is especially suited for the
analysis of multidimensional spectra. Applied re, the pleckstrin homo
logy domain, ARIA generated structures of good duality, and of suffici
ently high accuracy to solve the X-ray crystal structure of the same d
omain by molecular replacement. The comparison of the free NMR solutio
n structure to the X-ray structure, which is complexed to D-myo-inosit
ol-1,4,5-triphosphate, shows that the ligand primarily induces a disor
der-order transition in the binding loops, which are disordered in the
NMR ensemble but well ordered in the crystal. The structural core of
the protein is unaffected, as evidenced by a backbone root-mean-square
difference between the average NMR coordinates and the X-ray crystal
structure for the secondary structure elements of less than 0.6 Angstr
om. (C) 1997 Academic Press Limited.