caa a22 4500 477136613 CHVBK 20180405111720.0 cr unu---uuuuu 170330e19970201xx s 000 0 eng 10.1023/A:1018602220061 doi (NATIONALLICENCE)springer-10.1023/A:1018602220061 Automated probabilistic method for assigning backbone resonances of (13C,15N)-labeled proteins [Elektronische Daten] [Jonathan Lukin, Andrew Gove, Sarosh Talukdar, Chien Ho] We present a computer algorithm for the automated assignment of polypeptide backbone and13Cβ resonances of a protein of known primary sequence. Input to the algorithm consistsof cross peaks from several 3D NMR experiments: HNCA, HN(CA)CO, HN(CA)HA,HNCACB, COCAH, HCA(CO)N, HNCO, HN(CO)CA, HN(COCA)HA, and CBCA(CO)NH.Data from these experiments performed on glutamine-binding protein are analyzed statisticallyusing Bayes' theorem to yield objective probability scoring functions for matching chemicalshifts. Such scoring is used in the first stage of the algorithm to combine cross peaks fromthe first five experiments to form intraresidue segments of chemical shifts{Ni,HiN,Ciα,Ciβ,C′i}, while the latter five are combined into interresiduesegments {Ciα,Ciβ,C′i,Ni+1,HNi+1}. Given a tentative assignment of segments,the second stage of the procedure calculates probability scores based on the likelihood ofmatching the chemical shifts of each segment with (i) overlapping segments; and (ii) chemicalshift distributions of the underlying amino acid type (and secondary structure, if known). Thisjoint probability is maximized by rearranging segments using a simulated annealing program,optimized for efficiency. The automated assignment program was tested using CBCANH andCBCA(CO)NH cross peaks of the two previously assigned proteins, calmodulin and CheA.The agreement between the results of our method and the published assignments wasexcellent. Our algorithm was also applied to the observed cross peaks of glutamine-bindingprotein of Escherichia coli, yielding an assignment in excellent agreement with that obtainedby time-consuming, manual methods. The chemical shift assignment procedure described hereshould be most useful for NMR studies of large proteins, which are now feasible with the useof pulsed-field gradients and random partial deuteration of samples. Kluwer Academic Publishers, 1997 Three-dimensional NMR nationallicence Automated polypeptide resonance assignment nationallicence Lukin Jonathan Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, 15213-2683, Pittsburgh, PA, U.S.A aut Gove Andrew Robotics Institute, Carnegie Mellon University, 4400 Fifth Avenue, 15213-2683, Pittsburgh, PA, U.S.A aut Talukdar Sarosh Robotics Institute, Carnegie Mellon University, 4400 Fifth Avenue, 15213-2683, Pittsburgh, PA, U.S.A aut Ho Chien Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, 15213-2683, Pittsburgh, PA, U.S.A aut Journal of Biomolecular NMR Kluwer Academic Publishers 9/2(1997-02-01), 151-166 0925-2738 9:2<151 1997 9 10858 https://doi.org/10.1023/A:1018602220061 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1018602220061 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lukin Jonathan Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, 15213-2683, Pittsburgh, PA, U.S.A aut NATIONALLICENCE 700 1- Gove Andrew Robotics Institute, Carnegie Mellon University, 4400 Fifth Avenue, 15213-2683, Pittsburgh, PA, U.S.A aut NATIONALLICENCE 700 1- Talukdar Sarosh Robotics Institute, Carnegie Mellon University, 4400 Fifth Avenue, 15213-2683, Pittsburgh, PA, U.S.A aut NATIONALLICENCE 700 1- Ho Chien Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, 15213-2683, Pittsburgh, PA, U.S.A aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Kluwer Academic Publishers 9/2(1997-02-01), 151-166 0925-2738 9:2<151 1997 9 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer