Details

Autor(en) / Beteiligte

• Rose, Judith
• Müller, Boje
• Groscurth, Sira
• Giese, Jonas
• Eirich, Jürgen
• Finkemeier, Iris
• Twyman, Richard M.
• Prüfer, Dirk
• Noll, Gundula A.

Titel

The functionality of plant mechanoproteins (forisomes) is dependent on the dual role of conserved cysteine residues

Ist Teil von

• International journal of biological macromolecules, 2021-12, Vol.193 (Pt B), p.1332-1339

Ort / Verlag

Netherlands: Elsevier B.V

Erscheinungsjahr

2021

Quelle

MEDLINE

Beschreibungen/Notizen

• Forisomes are giant polyprotein complexes that undergo reversible conformational rearrangements from a spindle-like to a plug-like state in response to Ca2+ or changes in pH. They act as valves in the plant vasculature, and reproduce this function in vitro to regulate flow in microfluidic capillaries controlled by electro-titration. Heterologous expression in yeast or plants allows the large-scale production of tailor-made artificial forisomes for technical applications. Here we investigated the unexpected disintegration of artificial forisomes in response to Ca2+ following the deletion of the M1 motif in the MtSEO-F1 protein or the replacement of all four conserved cysteine residues therein. This phenomenon could be mimicked in wild-type forisomes under reducing conditions by adding a thiol alkylating agent. We propose a model in which reversible changes in forisome structure depend on cysteine residues with ambiguous redox states, allowing the formation of intermolecular disulfide bridges (confirmed by mass spectrometry) as well as noncovalent thiol interactions to connect forisome substructures in the dispersed state. This is facilitated by the projection of the M1 motif from the MtSEO-F1 protein as part of an extended loop. Our findings support the rational engineering of disintegrating forisomes to control the release of peptides or enzymes in microfluidic systems. •Deletion of conserved M1 motif leads to calcium-triggered forisome disintegration.•Conserved cysteines in the M1 motif preserve forisome functionality.•Disulfides and reduced cysteines are equally important for forisome integrity.