A protease activity displaying some thrombin-like characteristics in conditioned medium of zinnia mesophyll cells undergoing tracheary element differentiation

Xiao Hong Yu, Brian Jones, Alan M. Jones, Yair M. Heimer

    Research output: Contribution to journalArticlepeer-review

    2 Scopus citations

    Abstract

    The normal development of tracheary elements (TE) requires a selective degradation of the cytoplasm without loss of the extracellular wall that remains behind as the water-conducting units of xylem. Using zinnia-(Zinnia elegans L. cv. Green Envy) cultured mesophyll cells that synchronously transdifferentiate into TEs, extracellular and intracellular proteases, respectively, have been shown to both trigger death and to execute autolysis as the final component of a programmed cell death (PCD). We report here the appearance in the medium of an unusual proteolytic activity correlated with the PCD process just prior to the autolysis. The activity has a pH optimum of 5.5-6.0 and displays some thrombin characteristics. This protease activity has 1) a 10-fold higher affinity towards a thrombin-specific chromogenic substrate than toward a trypsin-specific chromogenic substrate; 2) a 1000-fold lower sensitivity to soybean trypsin inhibitor (STI) compared to trypsin; and 3) limited ability to cleave the protease-activated receptor-1, the native thrombin substrate. However, the addition of partially purified fraction containing the thrombin-like protease activity to the medium of PCD-competent cells does not prematurely trigger PCD, and the thrombin-specific peptide inhibitor phenylalanine-proline-aspartic acid-chloromethylketone fails to inhibit PCD or tracheary element (TE) formation. This suggests that this protease activity may play a role within the cells in execution of the autolysis or in the collapse of the tonoplast rather than as an extracellular proteolytic activity participating in the chain of events leading to cell death.

    Original languageEnglish
    Pages (from-to)292-300
    Number of pages9
    JournalJournal of Plant Growth Regulation
    Volume23
    Issue number4
    DOIs
    StatePublished - 1 Dec 2004

    Keywords

    • Programmed cell death
    • Thrombin
    • Tracheary elements
    • Zinnia

    ASJC Scopus subject areas

    • Agronomy and Crop Science
    • Plant Science

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