caa a22 4500 477136133 CHVBK 20180405111718.0 cr unu---uuuuu 170330e19970901xx s 000 0 eng 10.1023/A:1018350004729 doi (NATIONALLICENCE)springer-10.1023/A:1018350004729 Solution structures of staphylococcal nuclease from multidimensional, multinuclear NMR: Nuclease-H124L and its ternary complex with Ca2+ and thymidine-3′,5′-bisphosphate [Elektronische Daten] [Jinfeng Wang, Dagmar Truckses, Frits Abildgaard, Željko Džakula, Zsolt Zolnai, John Markley] The solution structures of staphylococcal nuclease (nuclease) H124L and itsternary complex, (nuclease-H124L)•pdTp•Ca2+, were determinedby ab initio dynamic simulated annealing using 1925 NOE, 119 φ, 20χ1 and 112 hydrogen bond constraints for the free protein,and 2003 NOE, 118 φ, 20 χ1 and 114 hydrogen bondconstraints for the ternary complex. In both cases, the final structuresdisplay only small deviations from idealized covalent geometry. In structuredregions, the overall root-mean-square deviations from mean atomic coordinatesare 0.46 (±0.05) Å and 0.41 (±0.05) Å for thebackbone heavy atoms of nuclease and its ternary complex, respectively. Thebackbone conformations of residues in the loop formed byArg81-Gly86, which is adjacent to the activesite, are more precisely defined in the ternary complex than in unligatednuclease. Also, the protein side chains that show NOEs and evidence forhydrogen bonds to pdTp (Arg35, Lys84,Tyr85, Arg87, Tyr113, andTyr115) are better defined in the ternary complex. As has beenobserved previously in the X-ray structures of nuclease-WT, the binding ofpdTp causes the backbone of Tyr113 to change from an extendedto a left-handed α-helical conformation. The NMR structures reportedhere were compared with available X-ray structures: nuclease-H124L [Truckseset al. (1996) Protein Sci., 5, 1907-1916] and the ternary complex ofwild-type staphylococcal nuclease [Loll and Lattman (1989) Proteins Struct.Funct. Genet., 5, 183-201]. Overall, the solution structures ofnuclease-H124L are consistent with these crystal structures, but smalldifferences were observed between the structures in the solution and crystalenvironments. These included differences in the conformations of certain sidechains, a reduction in the extent of helix 1 in solution, and many fewerhydrogen bonds involving side chains in solution. Kluwer Academic Publishers, 1997 Comparison of NMR and X-ray structures nationallicence Inhibitor binding nationallicence Stereospecific assignments nationallicence Distance constraints from NOEs nationallicence Torsion angle constraints from J-coupling nationallicence Hydrogen bond constraints nationallicence Structure refinement nationallicence Wang Jinfeng Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut Truckses Dagmar Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut Abildgaard Frits Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut Džakula Željko Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut Zolnai Zsolt Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut Markley John Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut Journal of Biomolecular NMR Kluwer Academic Publishers 10/2(1997-09-01), 143-164 0925-2738 10:2<143 1997 10 10858 https://doi.org/10.1023/A:1018350004729 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1018350004729 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wang Jinfeng Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut NATIONALLICENCE 700 1- Truckses Dagmar Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut NATIONALLICENCE 700 1- Abildgaard Frits Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut NATIONALLICENCE 700 1- Džakula Željko Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut NATIONALLICENCE 700 1- Zolnai Zsolt Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut NATIONALLICENCE 700 1- Markley John Department of Biochemistry, University of Wisconsin-Madison, 420 Henry Mall, 53706, Madison, WI, U.S.A aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Kluwer Academic Publishers 10/2(1997-09-01), 143-164 0925-2738 10:2<143 1997 10 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer