caa a22 4500 475744659 CHVBK 20180406123509.0 cr unu---uuuuu 170329e20000201xx s 000 0 eng 10.1023/A:1007034700573 doi (NATIONALLICENCE)springer-10.1023/A:1007034700573 Atomic Structures of Two Nitroxide Spin Labels Complexed with Human Thrombin: Comparison with Solution Studies [Elektronische Daten] [Vicki Nienaber, Lawrence Berliner] Crystal structures of thrombin complexed with two spin labels called para-V, 4-(2,2,5,5-tetramethyl-pyrrolidine-1-oxyl)-p-(fluorosulfonyl) benzamidine, and meta-V, 3-(2,2,5,5-tetramethyl-pyrrolidine-1-oxyl)-m-(fluorosulfonyl) benzamidine, have been completed at 2.0 and 3.0 Å resolution, respectively. Previous electron spin resonance studies with these labels gave rise to a low-resolution "topography map” of thrombin's extended active site. These labels monitor two distinct areas of the thrombin active site: (1) an apolar binding site which manifests itself in an biphasic activation/inhibition effect on thrombin activity and (2) a region sensitive to α-thrombin autoproteolytic cleavage(s) to γ-thrombin (Arg75-Tyr76 and/or Arg77A-Asn78, and Lys149E-Gly150, chymotrypsin numbering). Para-V was found to bind along the substrate binding cleft, while meta-V was found to bind both at the substrate primary specificity pocket and at a site which interacts with the γ-cleavage loop. These studies reaffirm that accurate information may be gained from solution studies and indicates the complementarity of solid-state studies. Plenum Publishing Corporation, 2000 Thrombin nationallicence spin labels nationallicence X-ray crystallography nationallicence electron spin resonance nationallicence serine protease nationallicence Nienaber Vicki Department of Chemical and Physical Sciences, The DuPont Merck Pharmaceutical Company, Experimental Station, 19880, Wilmington, Delaware aut Berliner Lawrence Department of Chemical and Physical Sciences, The DuPont Merck Pharmaceutical Company, Experimental Station, 19880, Wilmington, Delaware aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/2(2000-02-01), 129-137 0277-8033 19:2<129 2000 19 10930 https://doi.org/10.1023/A:1007034700573 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1007034700573 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nienaber Vicki Department of Chemical and Physical Sciences, The DuPont Merck Pharmaceutical Company, Experimental Station, 19880, Wilmington, Delaware aut NATIONALLICENCE 700 1- Berliner Lawrence Department of Chemical and Physical Sciences, The DuPont Merck Pharmaceutical Company, Experimental Station, 19880, Wilmington, Delaware aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/2(2000-02-01), 129-137 0277-8033 19:2<129 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer