Plant Physiol, February 2001, Vol. 125, pp. 835-846
Cabbage Cryoprotectin Is a Member of the Nonspecific Plant Lipid
Transfer Protein Gene Family1
Dirk K.
Hincha,*
Birgit
Neukamm,2
Hany A.M.
Sror,3
Frank
Sieg,4
Wolfgang
Weckwarth,
Markus
Rückels,
Valérie
Lullien-Pellerin,
Werner
Schröder, and
Jürgen M.
Schmitt
Institut für Pflanzenphysiologie und Mikrobiologie, Freie
Universität, Königin Luise-Strasse 12-16, D-14195 Berlin,
Germany (D.K.H., B.N., H.A.M.S., F.S., W.W., J.M.S.); Max-Planck
Institut für Molekulare Pflanzenphysiologie, D-14424 Potsdam,
Germany (D.K.H.); Boehringer Mannheim GmbH, Abteilung BP-Z 6, Division Biochemica, Nonnenwald 2, D-82372 Penzberg, Germany
(M.R.); Unité de Biochimie et de Biologie Moléculaire des
Céréales, Institut National de la Recherche Agronomique, 2 Place Viala, F-34060 Montpellier cedex 01, France (V.L.-P.); and
Institut für Biochemie, SFB 344, Freie Universität,
Fabeckstrasse 36a, D-14195 Berlin, Germany (W.S.)
We have recently purified a protein (cryoprotectin) from the leaves
of cold-acclimated cabbage (Brassica oleracea) to
electrophoretic homogeneity, which protects thylakoids isolated from
the leaves of nonacclimated spinach (Spinacia oleracea)
from freeze-thaw damage. Sequencing of cryoprotectin showed the
presence of at least three isoforms of WAX9 proteins, which belong to
the class of nonspecific lipid transfer proteins. Antibodies raised
against two synthetic peptides derived from the WAX9 proteins
recognized a band of approximately 10 kD in western blots of crude
cryoprotectin preparations. This protein and the cryoprotective
activity could be precipitated from solution by the antiserum. We show
further that cryoprotectin is structurally and functionally different from WAX9 isolated from the surface wax of cabbage leaves. WAX9 has
lipid transfer activity for phosphatidylcholine, but no cryoprotective activity. Cryoprotectin, on the other hand, has cryoprotective, but no
lipid transfer activity. The cryoprotective activity of cryoprotectin
was strictly dependent on Ca2+ and Mn2+ and
could be inhibited by chelating agents, whereas the lipid transfer
activity of WAX9 was higher in the presence of
ethylenediaminetetraacetate than in the presence of Ca2+
and Mn2+.
1
This work was supported by grants to J.M.S. and
D.K.H. and a Heisenberg stipend to D.K.H. from the Deutsche
Forschungsgemeinschaft. H.A.M.S. is supported by a stipend from the
Egyptian government.
2
Present address: FG Mikrobiologie und Genetik,
Technische Universität, Gustav-Meyer-Allee 25, D-13355 Berlin, Germany.
3
Permanent address: Biochemistry Department,
Faculty of Agriculture, Ain-shams University, Cairo, Egypt.
4
Present address: Institut für Physiologie,
Medizinische Fakultät, Otto-von-Guericke Universität,
Leipziger Str. 44, D-39120 Magdeburg, Germany.
*
Corresponding author; e-mail Hincha{at}mpimp-golm.mpg.de; fax
49-331-567-8250.
© 2001 American Society of Plant Physiologists
