Identification and characterization of a prevacuolar compartment in stigmas of Nicotiana alata

Elizabeth A. Miller, Marcus C.S. Lee, Marilyn A. Anderson (Lead / Corresponding author)

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40 Citations (Scopus)


The stigmas of the ornamental tobacco plant Nicotiana alata accumulate large quantities of a series of 6-kD proteinase inhibitors (PIs) in the central vacuole that are derived from a 40-kD precursor protein, Na-PI. The sorting information that directs Na-PI to the vacuole is likely to reside in a C-terminal propeptide domain of 25 amino acids that forms an amphipathic α helix. Using cell fractionation techniques, we have examined transit of Na-PI through the endomem brane system and have identified a prevacuolar compartment that contains Na-PI with an intact targeting signal. In contrast, the targeting signal is not present on the predominant form of Na-PI in the vacuole. The prevacuolar compartment is marked by the presence of homologs of both the t-SNARE, PEP12p, and the putative vacuolar sorting receptor BP-80. Cross-linking and affinity precipitation studies revealed that Na-PI associates with BP-80 within this compart ment, providing in vivo evidence for the function of BP-80 as a sorting receptor for a protein with a C-terminal vacuolar targeting signal.

Original languageEnglish
Pages (from-to)1499-1508
Number of pages10
JournalPlant Cell
Issue number8
Publication statusPublished - Aug 1999

ASJC Scopus subject areas

  • Plant Science
  • Cell Biology


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