Elsevier

Biochimie

Volume 166, November 2019, Pages 184-193
Biochimie

Review
Recombinant cystatins in plants

https://doi.org/10.1016/j.biochi.2019.06.006Get rights and content

Highlights

  • •

    Plant cystatins show value for the control of Cys proteases in biological systems.

  • •

    We review recent advances on the use of these proteins for crop trait improvement.

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    We then discuss their potential in plant protection for pest and pathogen control.

  • •

    We also address the fine-tuning of their activity against specific protease targets.

Abstract

Dozens of studies have assessed the practical value of plant cystatins as ectopic inhibitors of Cys proteases in biological systems. The potential of these proteins in crop protection to control herbivorous pests and pathogens has been documented extensively over the past 25 years. Their usefulness to regulate endogenous Cys proteases in planta has also been considered recently, notably to implement novel traits of agronomic relevance in crops or to generate protease activity-depleted environments in plants or plant cells used as bioreactors for recombinant proteins. After a brief update on the basic structural characteristics of plant cystatins, we summarize recent advances on the use of these proteins in plant biotechnology. Attention is also paid to the molecular improvement of their structural properties for the improvement of their protease inhibitory effects or the fine-tuning of their biological target range.

Introduction

Protease inhibitors of the cystatin protein superfamily are ubiquitous in plants, where they play several roles including the inhibition of insect digestive cathepsins upon herbivory challenge and the regulation of Cys proteases during programmed cell death, leaf senescence or storage protein mobilization in reproductive organs [1,2]. First identified in rice seeds by Soichi Arai, Keiko Abe and co-workers in the late 1980's [[3], [4], [5]], plant cystatins have been the object of intense work over the past three decades. Recent reviews have discussed the roles of these proteins as regulators of Cys protease-dependent cellular and physiological processes in planta [[6], [7], [8]]. Other reviews have discussed their involvement in plant defense and their possible usefulness in plant protection [9,10]. After a brief update on the basic structural elements of plant cystatins, we here summarize recent advances towards the use of these proteins in plant biotechnology. We also pay attention to the molecular engineering of their structural and functional properties for the improvement of their protease inhibitory effects or the fine-tuning of their effective activity range in biological contexts.

Section snippets

The basic structural elements of plant cystatins

Plant cystatins are competitive inhibitors of papain-like C1A Cys proteases harbouring the conserved structural motifs of cystatin protein superfamily members, i.e. a Gln–X–Val–X–Gly motif in the central region (where X is any amino acid), a Pro–Trp (or Leu–Trp) dipeptide motif in the C-terminal region, and a conserved Gly residue in the N-terminal region [2,11]. Hypervariable, positively selected amino acids are found within –or close to– these conserved motifs, that strongly influence the

Various Cys protease targets for cystatins in plant systems

Growing knowledge about the roles and modes of action of cystatins in plants has led over the years to promising developments towards the use of these proteins in plant genetic improvement. Numerous studies have assessed the potential of recombinant cystatins as anti-digestive proteins to engineer pest or pathogen resistance in major crops. Other studies have assessed their potential as ectopic modulators of endogenous proteases to introduce novel traits of agronomical value, or, more recently,

Conclusions and perspectives

Numerous papers have reported on the structure, engineering and biotechnological uses of plant cystatins since the publication of our previous review on this protein family in Biochimie some ten years ago [2]. Studies over this period have further confirmed the striking structural and functional similarities between these proteins and their animal counterparts, despite a number of features specific to each group of proteins. Other studies have confirmed the practical potential of plant

Acknowledgements

This work was financially supported by a Discovery grant from the Natural Science and Engineering Research Council of Canada and a Team project grant from the Fonds Québécois de la Recherche sur la Nature et les Technologies to D.M.

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