LETTERS
600 nature materials | VOL 2 | SEPTEMBER 2003 | www.nature.com/naturematerials
E
merging technologies are creating increasing interest in smart
materials that may serve as actuators in micro- and
nanodevices
1–3
. Mechanically active polymers currently studied
include a variety of materials
4–9
. ATP-driven motor proteins, the
actuators of living cells
10
, possess promising characteristics
11–13
, but their
dependence on strictly defined chemical environments can be
disadvantagous
14
. Natural proteins that deform reversibly by entropic
mechanisms might serve as models for artificial contractile polypeptides
with useful functionality
15
, but they are rare
16
. Protein bodies from sieve
elements of higher plants
17–19
provide a novel example. sieve elements
form microfluidics systems for pressure-driven transport of photo-
assimilates throughout the plant
20–22
. Unique protein bodies in the
sieve elements of legumes act as cellular stopcocks, by undergoing a
Ca
2+
-dependent conformational switch in which they plug the sieve
element
23
. In living cells, this reaction is probably controlled by Ca
2+
-
transporters in the cell membrane
23
. Here we report the rapid,
reversible, anisotropic and ATP-independent contractility in these
protein bodies in vitro. Considering the unique biological function of
the legume ‘crystalloid’ protein bodies and their contractile properties,
we suggest to give them the distinctive name forisome (‘gate-body’;
from the Latin foris, the wing of a gate).
We scrutinized the Ca
2+
response in vitro, using forisomes isolated
from broad bean (Vicia faba) sieve elements. Individual sieve elements
were ripped open with fine glass pipettes in Ca
2+
-free medium (see
Methods).The forisome was then pressed with the pipette tip against the
opposite cell wall to enable it to stick to the pipette and be transferred to
Ca
2+
-free medium in test vessels that allowed microscopic observation.
In our first attempts, forisomes were heavily damaged during the
extraction, but still swelled and shrank isodiametrically in response to
media with or without Ca
2+
, respectively (Fig. 1a). Forisomes that had
retained their shape (the length varied between 18 and 34 μm, and
diameter between 2.2 and 4 μm) during isolation and therefore
appeared intact, did not only swell radially in response to Ca
2+
, but also
contracted longitudinally (Fig. 1b). Thus, the condensed form of
forisomes corresponds to a longitudinally expanded state, whereas
forisomes in dispersed conformation are longitudinally contracted.
In contraction, forisomes shortened by 29.4% ± 6.7% (means ± s.d.,
n = 15) and increased in diameter (measured across their centre) by
119.5% ± 47.9%. This corresponds to a more than threefold Ca
2+
-
induced volume increase. The response to Ca
2+
, which invariably
occurred in all of the more than 200 intact forisomes that we isolated
individually, was fully reversible by transfer to Ca
2+
-free solutions
ATP-independent contractile proteins
from plants
MICHAEL KNOBLAUCH*
1
, GUNDULA A. NOLL
1
, TORSTEN MÜLLER
2
, DIRK PRÜFER
3
,
INGRID SCHNEIDER-HÜTHER
1
, DÖRTE SCHARNER
1
, AART J. E. VAN BEL
1
AND WINFRIED S. PETERS
1,4
1
Institut für Allgemeine Botanik der Justus Liebig-Universität, Senckenbergstr. 17-21, D-35390 Gießen, Germany
2
Institut für Biologie, Humboldt Universität zu Berlin, Invalidenstr. 42, D-10115 Berlin, Germany
3
Fraunhofer Institut für Molekularbiologie und Angewandte Ökologie, Bereich Molekularbiologie, Auf dem Aberg 1, D-57392 Schmallenberg-Grafschaft, Germany
4
Arbeitskreis Kinematische Zellforschung, Biozentrum der Johann Wolfgang Goethe-Universität, Marie-Curie-Str. 9, D-60439 Frankfurt am Main, Germany
*e-mail: michael.knoblauch@bio.bot1.uni-giessen.de
Published online: 24 August 2003; doi:10.1038/nmat960
Figure 1 Response to Ca
2+
of forisomes isolated from Vicia faba sieve elements.
a, Isotropic swelling of a forisome that had been damaged mechanically during
isolation. The forisome is attached to the tip of a glass pipette (tip diameter 4 μm).
Swelling and shrinkage were induced reversibly by application of Ca
2+
(10 mM CaCl
2
,
50 mM KCl, 10 mM HEPES pH 7.3) or Ca
2+
-free medium (as before, with CaCl
2
replaced by 10 mM disodium EDTA), respectively, as indicated. b, Anisotropic Ca
2+
response with longitudinal contraction by about 30% in an intact forisome (solutions
as in a). The length of the expanded forisome (left) is 24 μm. A movie showing this
experiment is available in the Supplementary Information, Movie S1.
©2003 Nature Publishing Group