First published online July 18, 2002; 10.1104/pp.002360
Plant Physiol, August 2002, Vol. 129, pp. 1664-1673
In Vivo and in Vitro Phosphorylation of Membrane and Soluble
Forms of Soybean Nodule Sucrose Synthase1
Olga
Komina,2
You
Zhou,
Gautam
Sarath, and
Raymond
Chollet*
Department of Biochemistry (O.K., G.S., R.C.) and Center for
Biotechnology (Y.Z., G.S.), University of Nebraska, Lincoln, Nebraska
68588-0664
Sucrose synthase (SS) is a known phosphoserine
(SerP)-containing enzyme in a variety of plant "sink" organs,
including legume root nodules, where it is phosphorylated
primarily at Ser-11. Using immunofluorescence confocal microscopy, we
documented that part of the total SS (nodulin-100) pool in mature
soybean (Glycine max) nodules is apparently associated
with the plasma membrane in situ, and we report that this association
is very "tight," as evidenced by a variety of chemical and
enzymatic pretreatments of the isolated microsomal fraction. To
investigate the in situ and in planta phosphorylation state of the
membrane (m) and soluble (s) forms of nodule SS, three complementary
approaches were used. First, excised nodules were radiolabeled in situ
with [32P]Pi for subsequent analysis of phosphorylated m-
and s-SS; second, immunopurified s- and m-SS were used as substrate in
"on-bead" assays of phosphorylation by nodule
Ca2+-dependent protein kinase; and third, SS-Ser-11(P)
phosphopeptide-specific antibodies were developed and used. The
collective results provide convincing evidence that microsomal
nodulin-100 is phosphorylated in mature nodules, and that it is
hypophosphorylated relative to s-SS (on an equivalent SS protein basis)
in attached, unstressed nodules. Moreover, the immunological data and
related phosphopeptide mapping analyses indicate that a homologous
N-terminal seryl-phosphorylation domain and site reside in microsomal
nodulin-100. We also observed that mild, short-term inorganic
nitrogen and salt stresses have a significant negative impact on
the content and N-terminal phosphorylation state of nodule m- and s-SS,
with the former being the more sensitive of the two SS forms.
1
This work was supported in part by the U.S.
National Science Foundation (grant no. MCB-9727236 to R.C.). This is
no. 13,595 in the University of Nebraska Agricultural Research Division
journal series.
2
Present address: Department of Computer Science and
Engineering, University of Nebraska, Ferguson Hall, Lincoln, NE
68588-0115.
*
Corresponding author; e-mail RCHOLLET1{at}unl.edu; fax
402-472-7842.
© 2002 American Society of Plant Physiologists
This article has been cited by other articles:
|
|
|
|
 
H. D. Coleman, J. Yan, and S. D. Mansfield
Sucrose synthase affects carbon partitioning to increase cellulose production and altered cell wall ultrastructure
PNAS,
August 4, 2009;
106(31):
13118 - 13123.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Wienkoop, E. Larrainzar, M. Glinski, E. M. Gonzalez, C. Arrese-Igor, and W. Weckwerth
Absolute quantification of Medicago truncatula sucrose synthase isoforms and N-metabolism enzymes in symbiotic root nodules and the detection of novel nodule phosphoproteins by mass spectrometry
J. Exp. Bot.,
September 1, 2008;
59(12):
3307 - 3315.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Persia, G. Cai, C. Del Casino, C. Faleri, M. T.M. Willemse, and M. Cresti
Sucrose Synthase Is Associated with the Cell Wall of Tobacco Pollen Tubes
Plant Physiology,
August 1, 2008;
147(4):
1603 - 1618.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Ladrera, D. Marino, E. Larrainzar, E. M. Gonzalez, and C. Arrese-Igor
Reduced Carbon Availability to Bacteroids and Elevated Ureides in Nodules, But Not in Shoots, Are Involved in the Nitrogen Fixation Response to Early Drought in Soybean
Plant Physiology,
October 1, 2007;
145(2):
539 - 546.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. White, J. Prell, E. K. James, and P. Poole
Nutrient Sharing between Symbionts
Plant Physiology,
June 1, 2007;
144(2):
604 - 614.
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. C. Hardin, K. A. Duncan, and S. C. Huber
Determination of Structural Requirements and Probable Regulatory Effectors for Membrane Association of Maize Sucrose Synthase 1
Plant Physiology,
July 1, 2006;
141(3):
1106 - 1119.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Wienkoop and W. Weckwerth
Relative and absolute quantitative shotgun proteomics: targeting low-abundance proteins in Arabidopsis thaliana
J. Exp. Bot.,
April 1, 2006;
57(7):
1529 - 1535.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. K. Azad, Y. Sawa, T. Ishikawa, and H. Shibata
Phosphorylation of Plasma Membrane Aquaporin Regulates Temperature-Dependent Opening of Tulip Petals
Plant Cell Physiol.,
May 1, 2004;
45(5):
608 - 617.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. C. Hardin, H. Winter, and S. C. Huber
Phosphorylation of the Amino Terminus of Maize Sucrose Synthase in Relation to Membrane Association and Enzyme Activity
Plant Physiology,
April 1, 2004;
134(4):
1427 - 1438.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Konishi, Y. Ohmiya, and T. Hayashi
Evidence That Sucrose Loaded into the Phloem of a Poplar Leaf Is Used Directly by Sucrose Synthase Associated with Various {beta}-Glucan Synthases in the Stem
Plant Physiology,
March 1, 2004;
134(3):
1146 - 1152.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Wienkoop and G. Saalbach
Proteome Analysis. Novel Proteins Identified at the Peribacteroid Membrane from Lotus japonicus Root Nodules
Plant Physiology,
March 1, 2003;
131(3):
1080 - 1090.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Silvente, A. Camas, and M. Lara
Heterogeneity of sucrose synthase genes in bean (Phaseolus vulgaris L.): evidence for a nodule-enhanced sucrose synthase gene
J. Exp. Bot.,
February 1, 2003;
54(383):
749 - 755.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
