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J Biol Chem, Vol. 275, Issue 14, 10016-10022, April 7, 2000
From the Horticultural Sciences and Plant Molecular and Cellular
Biology Program, University of Florida, Gainesville, Florida
32611
The Delta pH pathway is one of two systems for
protein transport to the thylakoid lumen. It is a novel transport
system that requires only the thylakoidal
Precursors Bind to Specific Sites on Thylakoid Membranes
prior to Transport on the Delta pH Protein Translocation
System*
and
pH to power translocation.
Several substrates of the Delta pH pathway, including the intermediate precursor form of OE17 (iOE17) and the truncated precursor form of OE17
(tOE17), were shown to bind to the membrane in the absence of the pH
and be transported into the lumen when the pH was restored. Binding
occurred without energy or soluble factors, and efficient transport
from the bound state (~80-90%) required only the pH. Binding is
due to protein-protein interactions because protease pretreatment of
thylakoids destroyed their binding capability. Precursors are bound to
a specific site on the Delta pH pathway because binding was competed by
saturating amounts of Delta pH pathway precursor proteins, but not by a
Sec pathway precursor protein. These results suggested that precursor
tOE17 binds to components of the Delta pathway translocation machinery.
Hcf106 and Tha4 are two components of the Delta pH pathway machinery. Antibodies to Hcf106 or Tha4, when prebound to thylakoids, specifically inhibited precursor transport on the Delta pH pathway. However, only
Hcf106 antibodies reduced the level of precursor binding. These results
suggest that Hcf106 functions in early steps of the transport process.
*
This work was supported in part by National Institutes of
Health Grant R01 GM46951 (to K. C.). This manuscript is Florida Agricultural Experiment Station Journal Series No. R-07351.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Present address: Institute of Biological Chemistry, Washington
State University, Pullman, WA 99164.
§
To whom correspondence should be addressed: Horticultural Sciences
Dept., Fifield Hall, University of Florida, Gainesville, FL 32611. Tel.: (352) 392-4711, ext. 219; Fax: (352) 392-5653; E-mail:
kcline@ufl.edu.
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