Summary
1H, 15N and 13C resonance assignments are presented for the group II phospholipase A2 (PLA2) from Agkistrodon piscivorus piscivorus. The secondary structure of the enzyme has been inferred from an analysis of coupling constants, interproton distances, chemical shifts, and kinetics of amide exchange. Overall, the secondary structure of this PLA2 is similar to the crystal structure of the homologous group II human nonpancreatic secretory phospholipase [Scott, D.L., White, S.P., Browning, J.L., Rosa, J.J., Gelb, M.H. and Sigler, P.B. (1991) Science, 254, 1007–1010]. In the group I enzyme from porcine pancreas, the amino-terminal helix becomes fully ordered in the ternary complex of enzyme, lipid micelles and inhibitor. The formation of this helix is thought to be important for the increase in activity of phospholipases on aggregated substrates [Van den Berg, B., Tessari, M., Boelens, R., Dijkman, R., De Haas, G.H., Kaptein, R. and Verheij, H.M. (1995) Nature Struct. Biol., 2, 402–406]. However, the group II enzyme from Agkistrodon piscivorus piscivorus possesses a defined and well-positioned aminoterminal helix in the absence of substrate. Therefore, there is a clear difference between the conformations of group I and group II enzymes in solution. These conformational differences suggest that formation of the amino-terminal helix is a necessary, but not sufficient, step in interfacial activation of phospholipases.
Similar content being viewed by others
Abbreviations
- PLA2:
-
phospholipase A2
- App-D49:
-
phospholipase from Agkistrodon piscivorus piscivorus
- NOE:
-
nuclear Overhauser effect
References
Almeida, P.F., Jerala, R., Rule, G.S. and Biltonen, R.L. (1995) Biophys. J., 68, A184.
Archer, S.J., Ikura, M., Torchia, D.A. and Bax, A. (1991) J. Magn. Reson., 95, 636–641.
Bax, A., Ikura, M., Kay, L.E., Torchia, D.A. and Tschudin, R. (1990a) J. Magn. Reson., 36, 304–318.
Bax, A., Clore, M. and Gronenborn, A.M. (1990b) J. Magn. Reson., 88, 425–431.
Bax, A. and Pochapsky, S. (1992) J. Magn. Reson., 99, 638–643.
Bax, A., Vuister, G.W., Grzesiek, S., Delaglio, F., Wang, A.C., Tschudin, R. and Zhu, G. (1994) Methods Enzymol., 239, 79–105.
Bell, J.D. and Biltonen, R.L. (1989) J. Biol. Chem., 264, 12194–12200.
Bell, J.D. and Biltonen, R.L. (1992) J. Biol. Chem., 267, 11046–11056.
Billeter, M., Braun, W. and Wüthrich, K. (1982) J. Mol. Biol., 155, 321–346.
Biltonen, R.L., Lathrop, B.K. and Bell, J.D. (1991) Methods Enzymol., 197, 234–248.
Bodenhausen, G. and Ruben, D.G. (1980) Chem. Phys. Lett., 69, 185–189.
Burack, W.R., Yuan, Q. and Biltonen, R.L. (1993) Biochemistry, 32, 583–589.
Burgoyne, R.D. and Morgan, A. (1990) Trends Biochem. Sci., 15, 365–366.
Cho, W., Tomaselli, A.G., Heinrikson, R.L. and Kezdy, F.J. (1988) J. Biol. Chem., 263, 11237–11241.
Clubb, R.T., Thanabal, V. and Wagner, G. (1992) J. Biomol. NMR, 2, 203–210.
Dalgarno, D.C., Levine, A. and Williams, R.J.P. (1983) Biosci. Rep., 3, 443–452.
Dennis, E.A. (1987) Biotechnology, 5, 1294–1300.
Driscoll, P.C., Clore, G.M., Marion, D., Wingfield, P.T. and Gronenborn, A.M. (1990) Biochemistry, 29, 3542–3556.
Dupureur, C.M., Yu, B.-Z., Jain, M.K., Noel, J.P., Deng, T., Li, Y., Byeon, I.-J.L. and Tsai, M.-D. (1992) Biochemistry, 32, 6402–6413.
Edison, A.S., Abilgaard, F., Westler, W.M., Mooberry, S. and Markley, J.L. (1994) Methods Enzymol., 239, 3–79.
Frenkiel, T., Bauer, C., Carr, M.D., Birdsall, B. and Feeney, J. (1990) J. Magn. Reson., 90, 420–425.
Grzesiek, S. and Bax, A. (1992a) J. Magn. Reson., 96, 432–440.
Grzesiek, S. and Bax, A. (1992b) J. Am. Chem. Soc., 114, 6291–6293.
Grzesiek, S. and Bax, A. (1993) J. Biomol. NMR, 3, 185–204.
Heinrikson, R.L., Krueger, E.T. and Keim, P.S. (1977) J. Biol. Chem., 252, 4913–4921.
Ikura, M., Bax, A., Clore, G.M. and Gronenborn, A.M. (1990) J. Am. Chem. Soc., 112, 9020–9022.
Jain, M.K. and Maliwal, B.P. (1993) Biochemistry, 32, 11838–11846.
Jerala, R. and Rule, G.S. (1995) J. Magn. Reson. Ser. B, 108, 294–298.
Kay, L.E., Ikura, M., Tschudin, R. and Bax, A. (1990) J. Magn. Reson., 89, 496–514.
Kuipers, O.P., Thunnissen, M.M.G.M., De, Geus, P., Dijkstra, B.W., Drenth, J., Verheij, H.M. and De, Haas, G.H. (1989) Science, 244, 82–85.
Lathrop, B.K., Burack, W.R., Biltonen, R.L. and Rule, G.S. (1992) Protein Exp. Purif., 3, 512–517.
Lichtenberg, D., Romero, G., Menashe, M. and Biltonen, R.L. (1986) J. Biol. Chem., 261, 5334–5340.
Maragonore, J.M. and Heinrikson, R.L. (1993) J. Biol. Chem., 268, 6064.
Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989a) J. Magn. Reson., 85, 393–399.
Marion, D., Ikura, M. and Bax, A. (1989b) J. Magn. Reson., 85, 425–430.
Menashe, M., Romero, G., Biltonen, R.L. and Lichtenberg, D. (1986) J. Biol. Chem., 261, 5328–5333.
Messerle, B.A., Wider, G., Otting, G., Weber, C. and Wüthrich, K. (1989) J. Magn. Reson., 85, 608–613.
Olejniczak, E.T. and Eaton, H. (1990) J. Magn. Reson., 87, 628–632.
Peters, A.P., Dekker, N., Van den, Berg, L., Boelens, R., Kaptein, R., Slotboom, A.J. and De, Haas, G. (1992) Biochemistry, 31, 10024–10030.
Renetseder, R., Brunie, S., Dijkstra, B.W., Drenth, J. and Sigler, P. (1985) J. Biol. Chem., 260, 11627–11634.
Scott, D.L., White, S.P., Otwinowski, Z., Yuan, W., Gelb, M.H. and Sigler, P.B. (1990) Science, 250, 1541–1546.
Scott, D.L., White, S.P., Browning, J.L., Rosa, J.J., Gelb, M.H. and Sigler, P.B. (1991) Science, 254, 1007–1010.
Scott, D.L. and Sigler, P.B. (1994) Adv. Protein Chem., 45, 53–88.
Slotboom, A.J., Verheij, H.M. and De, Haas, G.D. (1982) In Phospholipids (Eds, Hawthorne, J.N. and Ansell, G.B.), Elsevier Press, Amsterdam, The Netherlands.
Van, Dam-Mieras, M.C., Slotboom, A.J., Pieterson, W.A. and De, Haas, G.H. (1975) Biochemistry, 14, 5387–5394.
Van den, Berg, B., Tessari, M., Boelens, R., Dijkman, R., Kaptein, R. and De, Haas, G. (1995a) J. Biomol. NMR, 5, 110–121.
Van den, Berg, B., Tessari, M., Boelens, R., Dijkman, R., De, Haas, G., Kaptein, R. and Verheij, H.M. (1995b) Nature Struct. Biol., 2, 402–406.
Verheij, H.M., Egmond, M.R. and De, Haas, G.H. (1981) Biochemistry, 20, 94–99.
Vuister, G.W. and Bax, A. (1993) J. Am. Chem. Soc., 115, 7772–7777.
Wang, A.C., Lodi, P.J., Qin, J., Vuister, G.W., Gronenborn, A.M., and Clore, G.M. (1994) J. Magn. Reson. Ser. B, 105, 196–198.
Wishart, D.S. and Sykes, B.D. (1994) Methods Enzymol., 239, 363–392.
Wittekind, W. and Mueller, L. (1993) J. Magn. Reson. Ser. B, 101, 201–205.
Wüthrich, K. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York, NY.
Yamazaki, T., Forman-Kay, J.D. and Kay, L.E. (1993) J. Am. Chem. Soc., 115, 11054–11055.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Jerala, R., Almeida, P.F.F., Ye, Q. et al. 1H, 15N and 13C resonance assignments and secondary structure of group II phospholipase A2 from Agkistrodon piscivorus piscivorus: Presence of an amino-terminal helix in solution. J Biomol NMR 7, 107–120 (1996). https://doi.org/10.1007/BF00203821
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00203821