Zofie Sovová

Publications (4)7.86 Total impact

• Source

  Available from: Vladimír Kopecký

  Article: Structural analysis of natural killer cell receptor protein 1 (NKR-P1) extracellular domains suggests a conserved long loop region involved in ligand specificity.

  Zofie Sovová, Vladimír Kopecký, Tomáš Pazderka, Kateřina Hofbauerová, Daniel Rozbeský, Ondřej Vaněk, Karel Bezouška, Rüdiger Ettrich
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  ABSTRACT: Receptor proteins at the cell surface regulate the ability of natural killer cells to recognize and kill a variety of aberrant target cells. The structural features determining the function of natural killer receptor proteins 1 (NKR-P1s) are largely unknown. In the present work, refined homology models are generated for the C-type lectin-like extracellular domains of rat NKR-P1A and NKR-P1B, mouse NKR-P1A, NKR-P1C, NKR-P1F, and NKR-P1G, and human NKR-P1 receptors. Experimental data on secondary structure, tertiary interactions, and thermal transitions are acquired for four of the proteins using Raman and infrared spectroscopy. The experimental and modeling results are in agreement with respect to the overall structures of the NKR-P1 receptor domains, while suggesting functionally significant local differences among species and isoforms. Two sequence regions that are conserved in all analyzed NKR-P1 receptors do not correspond to conserved structural elements as might be expected, but are represented by loop regions, one of which is arranged differently in the constructed models. This region displays high flexibility but is anchored by conserved sequences, suggesting that its position relative to the rest of the domain might be variable. This loop may contribute to ligand-binding specificity via a coupled conformational transition.
  Journal of Molecular Modeling 06/2011; 17(6):1353-70. · 1.80 Impact Factor
• Source

  Available from: Vladimír Kopecký

  Article: Structure and dynamics of the N-terminal loop of PsbQ from photosystem II of Spinacia oleracea.

  Jaroslava Ristvejová, Vladimír Kopecký, Zofie Sovová, Mónica Balsera, Juan B Arellano, Michael Green, Rüdiger Ettrich
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  ABSTRACT: Infrared and Raman spectroscopy were applied to identify restraints for the structure determination of the 20 amino acid loop between two beta-sheets of the N-terminal region of the PsbQ protein of the oxygen evolving complex of photosystem II from Spinacia oleracea by restraint-based homology modeling. One of the initial models has shown a stable fold of the loop in a 20 ns molecular dynamics simulation that is in accordance with spectroscopic data. Cleavage of the first 12 amino acids leads to a permanent drift in the root means square deviation of the protein backbone and induces major structural changes.
  Biochemical and Biophysical Research Communications 07/2006; 345(1):287-91. · 2.48 Impact Factor
• Article: The phosphatase activity of the isolated H4-H5 loop of Na+/K+ ATPase resides outside its ATP binding site.

  Rita Krumscheid, Rüdiger Ettrich, Zofie Sovová, Klára Susánková, Zdenek Lánský, Katerina Hofbauerová, Holger Linnertz, Jan Teisinger, Evzen Amler, Wilhelm Schoner
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  ABSTRACT: The structural stability of the large cytoplasmic domain (H(4)-H(5) loop) of mouse alpha(1) subunit of Na(+)/K(+) ATPase (L354-I777), the number and the location of its binding sites for 2'-3'-O-(trinitrophenyl) adenosine 5'-triphosphate (TNP-ATP) and p-nitrophenylphosphate (pNPP) were investigated. C- and N-terminal shortening revealed that neither part of the phosphorylation (P)-domain are necessary for TNP-ATP binding. There is no indication of a second ATP site on the P-domain of the isolated loop, even though others reported previously of its existence by TNP-N(3)ADP affinity labeling of the full enzyme. Fluorescein isothiocyanate (FITC)-anisotropy measurements reveal a considerable stability of the nucleotide (N)-domain suggesting that it may not undergo a substantial conformational change upon ATP binding. The FITC modified loop showed only slightly diminished phosphatase activity, most likely due to a pNPP site on the N-domain around N398 whose mutation to D reduced the phosphatase activity by 50%. The amino acids forming this pNPP site (M384, L414, W411, S400, S408) are conserved in the alpha(1-4) isoforms of Na(+)/K(+) ATPase, whereas N398 is only conserved in the vertebrates' alpha(1) subunit. The phosphatase activity of the isolated H(4)-H(5) loop was neither inhibited by ATP, nor affected by mutation of D369, which is phosphorylated in native Na(+)/K(+) ATPase.
  European Journal of Biochemistry 11/2004; 271(19):3923-36. · 3.58 Impact Factor
• Article: The phosphatase activity of the isolated H4‐H5 loop of Na+/K+ ATPase resides outside its ATP binding site

  Rita Krumscheid, Rüdiger Ettrich, Zofie Sovová, Klára Sušánková, Zdeněk Lánský, Kateřina Hofbauerová, Holger Linnertz, Jan Teisinger, Evz̆en Amler, Wilhelm Schoner
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  ABSTRACT: The structural stability of the large cytoplasmic domain (H4-H5 loop) of mouse α1 subunit of Na+/K+ ATPase (L354–I777), the number and the location of its binding sites for 2′-3′-O-(trinitrophenyl) adenosine 5′-triphosphate (TNP-ATP) and p-nitrophenylphosphate (pNPP) were investigated. C- and N-terminal shortening revealed that neither part of the phosphorylation (P)-domain are necessary for TNP-ATP binding. There is no indication of a second ATP site on the P-domain of the isolated loop, even though others reported previously of its existence by TNP-N3ADP affinity labeling of the full enzyme. Fluorescein isothiocyanate (FITC)-anisotropy measurements reveal a considerable stability of the nucleotide (N)-domain suggesting that it may not undergo a substantial conformational change upon ATP binding. The FITC modified loop showed only slightly diminished phosphatase activity, most likely due to a pNPP site on the N-domain around N398 whose mutation to D reduced the phosphatase activity by 50%. The amino acids forming this pNPP site (M384, L414, W411, S400, S408) are conserved in the α1−4 isoforms of Na+/K+ ATPase, whereas N398 is only conserved in the vertebrates' α1 subunit. The phosphatase activity of the isolated H4-H5 loop was neither inhibited by ATP, nor affected by mutation of D369, which is phosphorylated in native Na+/K+ ATPase.
  European Journal of Biochemistry. 09/2004; 271(19):3923 - 3936.