Assignment strategy for proteins with known structure.
In protein NMR the assignment of nuclear spin resonances is a prerequisite for all subsequent applications, such as studies of ligand binding, protein-DNA interactions, and dynamics. Resonance assignment is a time consuming step even when the 3D x-ray structure of the protein is available. A new strategy is presented to solve the "inverse" assignment problem, which is the determination of the NMR resonance assignment from a known 3D protein structure. The protocol employs NMR data in the form of residual dipolar couplings and chemical shifts, while it does not require any sequential NMR connectivity information. The assignment problem is mathematically formulated in terms of a weighted matching problem that can be computationally efficiently solved by a combinatorial optimization algorithm. The protocol is applied to ubiquitin using two or three residual dipolar couplings per amino acid measured in Pfl phage medium together with chemical shift information. The algorithm yields for more than 90% of the protein backbone resonances the correct assignment.
Carlson School of Chemistry and Biochemistry, Clark University, Worcester, Massachusetts 01610, USA.
Jean-Christophe
JC
Hus
Jeanine J
JJ
Prompers
Rafael
R
Brüschweiler
D000465:Algorithms; D000595:Amino Acid Sequence; D008024:Ligands; D008962:Models, Theoretical; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry; D025801:Ubiquitin / Q000737:chemistry*
D008024:Ligands; D011506:Proteins; D025801:Ubiquitin