Characterizing the Na+/K+-ATPase

  • Joseph D. Robinson
Part of the People and Ideas Series book series (PEOPL)


By 1965 numerous approaches had converged, supporting proposals that (Nat+ K+)—stimulated ATPase activity underlay the Na+/K+ pump of higher animals.’ Later papers in vast profusion reported diverse studies of ATPase and pump, describing details of details, but also demonstrating unequivocally the identity of pump and ATPase. Here only a drastically condensed sketch can be achieved. My aim is to illustrate the extensive branching of experimental endeavors that revealed complexities in this entity—a multi-sited enzyme—and in the process—cation transport by multiple modes linked to particular sequences of enzymatic reaction steps.


ATPase Activity Transport Mode Occlude State Alternative Conformation ATPase Reaction 
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Notes to Chapter 11

  1. 1.
    Through discovery of alternative mechanisms and failure to find the Na+/K+ pump, it became clear that bacteria, fungi, plants, and lower animals do not possess the Na+/K+- ATPase described in molluscs, arthropods, and chordates.Google Scholar
  2. 2.
    Garrahan and Glynn (1967a), p. 160.Google Scholar
  3. 3.
    Garrahan and Glynn (1967e). These experiments showed incorporation of 32P from P, into ATP; later experiments showed net synthesis of ATP (Lew et al., 1970).Google Scholar
  4. 4.
    Garrahan and Glynn (1967a,c,d).Google Scholar
  5. 5.
    Glynn and Hoffman (1971).Google Scholar
  6. 6.
    Cavieres and Glynn (1979).Google Scholar
  7. 7.
    Glynn et al. (1971).Google Scholar
  8. 8.
    Simons (1974, 1975).Google Scholar
  9. 9.
    Garrahan and Glynn (1967a,b).Google Scholar
  10. 10.
    Lew et al. (1973); Glynn and Karlish (1976).Google Scholar
  11. 11.
    Lee and Blostein (1980); Blostein (1983).Google Scholar
  12. 12.
    Forgac and Chin (1982).Google Scholar
  13. 13.
    Sachs (1986).Google Scholar
  14. 14.
    Mullins and Brinley (1969).Google Scholar
  15. 15.
    For example, Robinson (1967, 1969) described cooperative binding of cations to the ATPase, following the allosteric models of Monod et al. (1963, 1965). Activation could then occur without all sites being filled, in contrast to models requiring that three Na+ and two K+ necessarily be bound.Google Scholar
  16. 16.
    Abercrombie and De Weer (1978); De Weer and Geduldig (1978); Rakowski et al. (1989).Google Scholar
  17. 17.
    Thomas (1969).Google Scholar
  18. 18.
    Uesugi et al. (1971).Google Scholar
  19. 19.
    However, further experience demonstrated that membrane-bound enzymes, which necessarily have extensive hydrophobic surfaces, may migrate differently from “soluble” proteins of equal molecular weight.Google Scholar
  20. 20.
    Kyte (1971a).Google Scholar
  21. 21.
    In photographs of Hokin’s gels, bands are visible not only at 94 kDa, but also at 64, 53, and 35 kDa.Google Scholar
  22. 22.
    Kyte (1971b, 1972).Google Scholar
  23. 23.
    Jorgensen and Skou (1969).Google Scholar
  24. 24.
    Jorgensen and Skou (1971); Jorgensen et al. (1971). By that time (and before Kyte’s paper appeared), Skou had detected two bands by SDS-PAGE, but after some disagreement, they chose not to describe the finding (Skou, interview, 1993).Google Scholar
  25. 25.
    Jorgensen (1974a,b).Google Scholar
  26. 26.
    Nakao et al. (1974); Lane (1973); Dixon and Hokin (1974); Hokin et al. (1973).Google Scholar
  27. 27.
    See, for example, Robinson and Flashner (1979); Jorgensen (1982); Glynn (1985).Google Scholar
  28. 28.
    See, for example, Repke and Schön (1973) and Askari et al. (1980) vs. Brotherus et al. (1981, 1983) and Jorgensen and Andersen (1986). A similar issue, whether two ATP-binding sites coexist on the enzyme, was strongly advocated by, among others, Askari (1987), Scheiner-Bobis et al. (1987), and Schuurmans-Stekhoven et al. (1981); however, Norby (1987) could find only one ATP-binding site per a-subunit.Google Scholar
  29. 29.
    A reasonable concern was whether subunits necessary for transport might be selectively lost during the purification procedure.Google Scholar
  30. 30.
    Kagawa and Racker (1971); Racker (1972, 1973). The phospholipid vesicles are often termed “liposomes.”Google Scholar
  31. 31.
    Hilden et al. (1974).Google Scholar
  32. 32.
    Goldin and Tong (1974). This paper was submitted (and appeared) two and a half months before the paper by Hilden et al.Google Scholar
  33. 33.
    Sweadner and Goldin (1975).Google Scholar
  34. 34.
    Hilden and Hokin (1975).Google Scholar
  35. 35.
    Goldin (1977).Google Scholar
  36. 36.
    Fahn et al. (1966a,b).Google Scholar
  37. 37.
    Fahn et al. (1966a), p. 1888.Google Scholar
  38. 38.
    Bader et al. (1966, 1967, 1968).Google Scholar
  39. 39.
    Post et al. (1969).Google Scholar
  40. 40.
    Kahlenberg et al. (1967, 1968).Google Scholar
  41. 41.
    Post and Kume (1973).Google Scholar
  42. 42.
    Degani et al. (1974); Hokin (1974). Degani et al. noted that their study was prompted by Post’s finding, and Hokin noted that his reinvestigation was prompted by Degani and Boyer’s development of their technique.Google Scholar
  43. 43.
    Albers (1967); Siegel and Albers (1967).Google Scholar
  44. 44.
    Albers (1967), p. 743.Google Scholar
  45. 45.
    This is not paradoxical since the schemes remembered are those having some general validity, and schemes are necessarily modified in detail as further studies reveal intricacies characteristic of biological processes.Google Scholar
  46. 46.
    Beaugé and Glynn (1979a).Google Scholar
  47. 47.
    Robinson (1974a,b); Klodos and Skou (1975).Google Scholar
  48. 48.
    Post et al. (1973, 1975).Google Scholar
  49. 49.
    Post et al. (1972).Google Scholar
  50. 50.
    Jorgensen (1975, 1977).Google Scholar
  51. 51.
    Karlish and Yates (1978).Google Scholar
  52. 52.
    Karlish (1980). He reported that only the a-peptide was labeled, with one to two FITC per peptide.Google Scholar
  53. 53.
    Skou and Esmann (1980, 1981).Google Scholar
  54. 54.
    Kapakos and Steinberg (1982, 1986a,b); Tyson et al. (1989).Google Scholar
  55. 55.
    Steinberg and Karlish (1989); Stürmer et al. (1989); Pratap et al. (1991).Google Scholar
  56. 56.
    Repke (1963); the meeting was held in 1961.Google Scholar
  57. 57.
    See, for example, Schwartz et al. (1969); Akera et al. (1969).Google Scholar
  58. 58.
    Repke (1964); Glynn (1964). This proposal was developed by Langer (1972), who included the Na+/Ca2+ exchanger linking Na+ and Ca2+ fluxes.Google Scholar
  59. 59.
    Schwartz et al. (1968). Charnock et al. (1963) had previously proposed that the cardiotonic steroids reacted with the phosphorylated enzyme.Google Scholar
  60. 60.
    Lindenmayer et al. (1968).Google Scholar
  61. 61.
    Albers et al. (1968).Google Scholar
  62. 62.
    Siegel et al. (1969); Sen et al. (1969); Post et al. (1969).Google Scholar
  63. 63.
    Ruoho and Kyte (1974).Google Scholar
  64. 64.
    Post et al. (1973, 1975).Google Scholar
  65. 65.
    Taniguchi and Post (1975).Google Scholar
  66. 66.
    Cantley and Josephson (1976); Josephson and Cantley (1977); Cantley et al. (1977, 1978). Vanadate was a previously unrecognized contaminant of ATP prepared from horse muscle.Google Scholar
  67. 67.
    Robinson et al. (1986). Arguments for the participation of Ala+ from glassware and for the resemblance of A1F4 - to phosphate were presented by Sternweis and Gilman (1982) and by Bigay et al. (1985).Google Scholar
  68. 68.
    Hegyvary and Post (1971); Nerby and Jensen (1971); Jensen and Nerby (1971).Google Scholar
  69. 69.
    Hegyvary and Post (1971), p. 5239.Google Scholar
  70. 70.
  71. 71.
    Kanazawa et al. (1970).Google Scholar
  72. 72.
    See, for example, Robinson (1976); Askari (1987).Google Scholar
  73. 73.
    Smith et al. (1980). Moczydlowski and Fortes (1981) made the same argument.Google Scholar
  74. 74.
    Post and Sen (1965); they attributed the notion of an intermediate stage to the analysis by Rosenberg and Wilbrandt (1957). Post and Sen repeated their proposal in 1969 (Post et al., 1969) with the argument that otherwise a leak could occur.Google Scholar
  75. 75.
    Post et al. (1972).Google Scholar
  76. 76.
    Post et al. selected Rb+ and Li+ empirically for their extreme effects. By that time Rb+ was known to be a good substitute for K+ in the Na+/K+-ATPase reaction and Li+ a poor substitute.Google Scholar
  77. 77.
    Karlish and Yates (1978).Google Scholar
  78. 78.
    Beaugé and Glynn (1979b).Google Scholar
  79. 79.
    K+ was thought to be binding from the cytoplasmic surface and from there moving to the occluded state. In the forward direction, K+ binds from the extracellular surface (when the enzyme is in the Ez-P state) and from there moves to the occluded state. Release from the occluded state would then be from the cytoplasmic surface.Google Scholar
  80. 80.
    Glynn and Richards (1982).Google Scholar
  81. 81.
    Forbush (1987).Google Scholar
  82. 82.
    Glynn et al. (1984).Google Scholar
  83. 83.
    Esmann and Skou (1985).Google Scholar
  84. 84.
    Karlish et al. (1990).Google Scholar
  85. 85.
    Biographical information is from interviews and correspondence (1993–1995).Google Scholar
  86. 86.
    Interview (1993); Skou (1989).Google Scholar
  87. 87.
    Interview (1993).Google Scholar

Copyright information

© American Physiological Society 1997

Authors and Affiliations

  • Joseph D. Robinson
    • 1
  1. 1.Department of PharmacologyState University of New York Health Science CenterSyracuseUSA

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