caa a22 4500 445325038 CHVBK 20180317142717.0 cr unu---uuuuu 170323e20110201xx s 000 0 eng 10.1007/s10858-010-9467-z doi (NATIONALLICENCE)springer-10.1007/s10858-010-9467-z RDC derived protein backbone resonance assignment using fragment assembly [Elektronische Daten] [Xingsheng Wang, Brian Tash, John Flanagan, Fang Tian] Experimental residual dipolar couplings (RDCs) in combination with structural models have the potential for accelerating the protein backbone resonance assignment process because RDCs can be measured accurately and interpreted quantitatively. However, this application has been limited due to the need for very high-resolution structural templates. Here, we introduce a new approach to resonance assignment based on optimal agreement between the experimental and calculated RDCs from a structural template that contains all assignable residues. To overcome the inherent computational complexity of such a global search, we have adopted an efficient two-stage search algorithm and included connectivity data from conventional assignment experiments. In the first stage, a list of strings of resonances (CA-links) is generated via exhaustive searches for short segments of sequentially connected residues in a protein (local templates), and then ranked by the agreement of the experimental 13Cα chemical shifts and 15N-1H RDCs to the predicted values for each local template. In the second stage, the top CA-links for different local templates in stage I are combinatorially connected to produce CA-links for all assignable residues. The resulting CA-links are ranked for resonance assignment according to their measured RDCs and predicted values from a tertiary structure. Since the final RDC ranking of CA-links includes all assignable residues and the assignment is derived from a "global minimum”, our approach is far less reliant on the quality of experimental data and structural templates. The present approach is validated with the assignments of several proteins, including a 42kDa maltose binding protein (MBP) using RDCs and structural templates of varying quality. Since backbone resonance assignment is an essential first step for most of biomolecular NMR applications and is often a bottleneck for large systems, we expect that this new approach will improve the efficiency of the assignment process for small and medium size proteins and will extend the size limits assignable by current methods for proteins with structural models. Springer Science+Business Media B.V., 2010 Backbone resonance assignment nationallicence Residual dipolar couplings nationallicence Structure-assisted resonance assignment nationallicence Wang Xingsheng Department of Biochemistry and Molecular Biology, College of Medicine, Pennsylvania State University, 17033, Hershey, PA, USA aut Tash Brian Department of Biochemistry and Molecular Biology, College of Medicine, Pennsylvania State University, 17033, Hershey, PA, USA aut Flanagan John Department of Biochemistry and Molecular Biology, College of Medicine, Pennsylvania State University, 17033, Hershey, PA, USA aut Tian Fang Department of Biochemistry and Molecular Biology, College of Medicine, Pennsylvania State University, 17033, Hershey, PA, USA aut Journal of Biomolecular NMR Springer Netherlands 49/2(2011-02-01), 85-98 0925-2738 49:2<85 2011 49 10858 https://doi.org/10.1007/s10858-010-9467-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-010-9467-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wang Xingsheng Department of Biochemistry and Molecular Biology, College of Medicine, Pennsylvania State University, 17033, Hershey, PA, USA aut NATIONALLICENCE 700 1- Tash Brian Department of Biochemistry and Molecular Biology, College of Medicine, Pennsylvania State University, 17033, Hershey, PA, USA aut NATIONALLICENCE 700 1- Flanagan John Department of Biochemistry and Molecular Biology, College of Medicine, Pennsylvania State University, 17033, Hershey, PA, USA aut NATIONALLICENCE 700 1- Tian Fang Department of Biochemistry and Molecular Biology, College of Medicine, Pennsylvania State University, 17033, Hershey, PA, USA aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 49/2(2011-02-01), 85-98 0925-2738 49:2<85 2011 49 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer