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Micromechanical measurements on P-protein aggregates (forisomes) from Vicia faba plants

 
: Schwan, S.; Menzel, M.; Fritzsche, M.; Heilmann, A.; Spohn, U.

:

Biophysical chemistry 139 (2009), Nr.2-3, S.99-105
ISSN: 0301-4622
Englisch
Zeitschriftenaufsatz
Fraunhofer IWM ()
Fraunhofer IWM-H ()
P-protein aggregate; chemo-mechanical activity; influence of metal ions and pH; force measurement

Abstract
Forisomes are chemomechanically active P-protein aggregates found in the phloem. of legumes. They can convert chemical energy into mechanical work when induced by divalent metal ions or changes in pH, which control the folding state of individual forisome proteins. We investigated the changing geometric parameters of individual forisomes and the strength and dynamics of the forces generated during this process. Three different divalent ions were tested (Ca2+, Sr2+ and Ba2+) and were shown to induce similar changes to the normalized length and diameter. In the concentration range from 0.1 to 4 M, K+ and Cl- ions had no influence on the contraction behaviour of the forisomes induced by 10 mM Ca2+. In the absence of dissolved oxygen, these changes were independent of the radius of the metal ion, water uptake and the strength of binding between the selected metal ions and those protein molecules responsible for forisome conformational transformation. In the absence of any load, bound Ca2+, Sr2+ and Ba2+ ions showed apparent and averaged dissociation constants of 14, 62 and 1070 mu M, respectively. Various forisomes generated bending on a quartz glass fibre with a diameter of 9 pm. The fibre bending was measured microscopically also by correlation between the digital patterns of a predefined window of observation before and after bending. Similar bending forces of approximately 90 nN were measured for a single forisome sequentially exposed to 10 mM Ca2+, Sr2+ and Ba2+. In the absence of dissolved oxygen, the same conditions resulted in averaged bending forces of (93 +/- 40) nN, (58 +/- 20) nN, and (91 +/- 20) nN after contacting different forisomes with 10 mM Ca2+, 10 mM Sr2+, and 10 mM Ba2+ respectively, demonstrating that the force generated was independent on ion concentrations above a certain threshold value.

: http://publica.fraunhofer.de/dokumente/N-91270.html