Cyanobacterial and chloroplast F1-ATPases: cross-reconstitution of photophosphorylation and subunit immunological relationships

David B. Hicks, Nathan Nelson, Charles F. Yocum

Research output: Contribution to journalArticlepeer-review

Abstract

The photosynthetic F1-ATPase from the cyanobacterium Spirulina platensis was recently purified in a five-subunit, reconstitutively active form (Hicks, D.B. and Yocum, C.F. (1986) Arch. Biochem. Biophys. 245, 220-229). Here we report on the similarities of the cyanobacterial F1 to the higher plant chloroplast F1 (CF1), as judged by two distinct methods. The ability of each coupling factor to reconstitute photophosphorylation in photosynthetic membranes depleted of F1 content by 2 M NaBr treatment was tested. Addition of either the homologous enzyme (e.g., Spirulina F1, Spirulina membranes) or the heterologous enzyme (e.g., spinach CF1 Spirulina membranes) to depleted membranes increased the rate of phenazine methosulfate-dependent cyclic photophosphorylation from nearly zero to up to 70 μmol ATP/h per mg Chl. Antibodies against four subunits of CF1 (α, γ, δ and ε) and against β of Escherichia coli F1 were reacted with the Spirulina enzyme by protein blotting. The α, β and γ subunits of Spirulina F1 cross-reacted with antibodies against the corresponding subunits from spinach. The cross-reactivity of the γ subunit correlated with previous observations that Spirulina membrane ATPase activity can be modulated by light and dithiothreitol, in a similar fashion to their effect on the enzyme from spinach chloroplasts. The ability of cyanobacterial and chloroplast enzymes to restore activity to heterologous membranes in the absence of immunological similarities between their respective δ and ε subunits may suggest that structural features other than particular amino acid sequences of these subunits are paramount in their roles in binding F1 to the membrane and in sealing proton leaks.

Original languageEnglish
Pages (from-to)217-222
Number of pages6
JournalBiochimica et Biophysica Acta - Bioenergetics
Volume851
Issue number2
DOIs
StatePublished - 10 Sep 1986
Externally publishedYes

Keywords

  • (Spirulina, Spinach, Chloroplast)
  • ATPase
  • Coupling factor
  • Photophosphorylation
  • Reconstitution

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