|
Mutagenesis of the Glucose-1-Phosphate-Binding Site of Potato
Tuber ADP-Glucose Pyrophosphorylase1
Yingbin Fu,
Miguel A. Ballicora, and
Jack Preiss*
Department of Biochemistry, Michigan State University, East
Lansing, Michigan 48824
Lysine (Lys)-195 in the
homotetrameric ADP-glucose pyrophosphorylase (ADPGlc PPase) from
Escherichia coli was shown previously to be involved in
the binding of the substrate glucose-1-phosphate (Glc-1-P). This
residue is highly conserved in the ADPGlc PPase family. Site-directed
mutagenesis was used to investigate the function of this conserved Lys
residue in the large and small subunits of the heterotetrameric potato
(Solanum tuberosum) tuber enzyme. The apparent affinity
for Glc-1-P of the wild-type enzyme decreased 135- to 550-fold by
changing Lys-198 of the small subunit to arginine, alanine, or glutamic
acid, suggesting that both the charge and the size of this residue
influence Glc-1-P binding. These mutations had little effect on the
kinetic constants for the other substrates (ATP and Mg2+ or
ADP-Glc and inorganic phosphate), activator (3-phosphoglycerate), inhibitor (inorganic phosphate), or on the thermal stability. Mutagenesis of the corresponding Lys (Lys-213) in the large subunit had
no effect on the apparent affinity for Glc-1-P by substitution with
arginine, alanine, or glutamic acid. A double mutant,
SK198RLK213R, was also obtained that had a
100-fold reduction of the apparent affinity for Glc-1-P. The data
indicate that Lys-198 in the small subunit is directly involved in the
binding of Glc-1-P, whereas they appear to exclude a direct role of
Lys-213 in the large subunit in the interaction with this substrate.
1
This work was supported in part by Department of
Energy grant no. DE-FG02-93ER20121.
*
Corresponding author; e-mail preiss{at}pilot.msu.edu; fax
1-517-353-9334.
Plant Physiol. (1998) 117: 989-996
Copyright Clearance Center: 0032-0889/98/117/0989/08
© 1998 American Society of Plant Physiologists
This article has been cited by other articles:
|
|
|
|
 
S.-K. Hwang, Y. Nagai, D. Kim, and T. W. Okita
Direct Appraisal of the Potato Tuber ADP-glucose Pyrophosphorylase Large Subunit in Enzyme Function by Study of a Novel Mutant Form
J. Biol. Chem.,
March 14, 2008;
283(11):
6640 - 6647.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Georgelis, E. L. Braun, J. R. Shaw, and L. C. Hannah
The Two AGPase Subunits Evolve at Different Rates in Angiosperms, yet They Are Equally Sensitive to Activity-Altering Amino Acid Changes When Expressed in Bacteria
PLANT CELL,
May 1, 2007;
19(5):
1458 - 1472.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. M. Bejar, X. Jin, M. A. Ballicora, and J. Preiss
Molecular Architecture of the Glucose 1-Phosphate Site in ADP-glucose Pyrophosphorylases
J. Biol. Chem.,
December 29, 2006;
281(52):
40473 - 40484.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. A. Iglesias, M. A. Ballicora, J. I. Sesma, and J. Preiss
Domain Swapping between a Cyanobacterial and a Plant Subunit ADP-Glucose Pyrophosphorylase
Plant Cell Physiol.,
April 1, 2006;
47(4):
523 - 530.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. A. Ballicora, J. R. Dubay, C. H. Devillers, and J. Preiss
Resurrecting the Ancestral Enzymatic Role of a Modulatory Subunit
J. Biol. Chem.,
March 18, 2005;
280(11):
10189 - 10195.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Crevillen, M. A. Ballicora, A. Merida, J. Preiss, and J. M. Romero
The Different Large Subunit Isoforms of Arabidopsis thaliana ADP-glucose Pyrophosphorylase Confer Distinct Kinetic and Regulatory Properties to the Heterotetrameric Enzyme
J. Biol. Chem.,
August 1, 2003;
278(31):
28508 - 28515.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. A. Ballicora, A. A. Iglesias, and J. Preiss
ADP-Glucose Pyrophosphorylase, a Regulatory Enzyme for Bacterial Glycogen Synthesis
Microbiol. Mol. Biol. Rev.,
June 1, 2003;
67(2):
213 - 225.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Koller, M. P. Washburn, B. M. Lange, N. L. Andon, C. Deciu, P. A. Haynes, L. Hays, D. Schieltz, R. Ulaszek, J. Wei, et al.
From the Cover: Proteomic survey of metabolic pathways in rice
PNAS,
September 3, 2002;
99(18):
11969 - 11974.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. B. Frueauf, M. A. Ballicora, and J. Preiss
Aspartate Residue 142 Is Important for Catalysis by ADP-glucose Pyrophosphorylase from Escherichia coli
J. Biol. Chem.,
November 30, 2001;
276(49):
46319 - 46325.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. A. Ballicora, J. B. Frueauf, Y. Fu, P. Schurmann, and J. Preiss
Activation of the Potato Tuber ADP-glucose Pyrophosphorylase by Thioredoxin
J. Biol. Chem.,
January 14, 2000;
275(2):
1315 - 1320.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Lal, J.-H. Choi, J. R. Shaw, and L. C. Hannah
A Splice Site Mutant of Maize Activates Cryptic Splice Sites, Elicits Intron Inclusion and Exon Exclusion, and Permits Branch Point Elucidation
Plant Physiology,
October 1, 1999;
121(2):
411 - 418.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
Y. Fu, M. A. Ballicora, J. F. Leykam, and J. Preiss
Mechanism of Reductive Activation of Potato Tuber ADP-glucose Pyrophosphorylase
J. Biol. Chem.,
September 25, 1998;
273(39):
25045 - 25052.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. A. Ballicora, Y. Fu, N. M. Nesbitt, and J. Preiss
ADP-Glucose Pyrophosphorylase from Potato Tubers. Site-Directed Mutagenesis Studies of the Regulatory Sites
Plant Physiology,
September 1, 1998;
118(1):
265 - 274.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
I. H. Kavakli, J.-S. Park, C. J. Slattery, P. R. Salamone, J. Frohlick, and T. W. Okita
Analysis of Allosteric Effector Binding Sites of Potato ADP-glucose Pyrophosphorylase through Reverse Genetics
J. Biol. Chem.,
October 26, 2001;
276(44):
40834 - 40840.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
