A helical element in the C-terminal domain of the N. plumbaginifolia F1 beta presequence is important for recognition by the mitochondrial processing peptidase.
The mitochondrial processing peptidase (MPP) in lower eucaryots and mammals is a matrix enzyme, whereas MPP in plants constitutes an integral part of the bc1 complex of the respiratory chain. The isolated spinach leaf bc1 complex catalyzes cleavage of the precursor of Nicotiana plumbaginifolia F1 beta subunit of the ATP synthase, resulting in a production of mature protein and a presequence that consists of 54 amino acids. A synthetic peptide derived from the C-terminal part of the presequence, containing 17 amino acids with a helical structural element, p-F1 beta(38-54), was an efficient inhibitor of the processing, whereas a peptide derived from the N-terminal part of the presequence, p-F1 beta(1-18), was much less effective. ATIII, a helical peptide derived from antithrombin III, was not recognized by MPP. Synthetic peptides corresponding to 4, 6, and 11 amino acids of the C terminus of the presequence, p-F1 beta(51-54), p-F1 beta(49-54), and p-F1 beta(44-54) were almost completely inert. Competition studies show that MPP recognizes the C-terminal domain of the presequence rather than the N-terminal domain. Furthermore, the alpha-helical element of the C-terminal domain is shown to be required for the recognition event.
Sjöling
S
S
Department of Biochemistry, Arrhenius Laboratories for Natural Sciences, Stockholm University, Sweden.
Eriksson
A C
AC
Glaser
E
E
D000595:Amino Acid Sequence; D008666:Metalloendopeptidases / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D011499:Protein Processing, Post-Translational; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000378:metabolism*; D018724:Spinacia oleracea / Q000201:enzymology; D013379:Substrate Specificity; D014026:Nicotiana / Q000378:metabolism*; D000096122:Mitochondrial Processing Peptidase
D008666:Metalloendopeptidases; D006180:Proton-Translocating ATPases; D000096122:Mitochondrial Processing Peptidase