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Generation, characterization, and heterologous expression of wild-type and up-regulated forms of Arabidopsis thaliana leaf ADP-glucose pyrophosphorylase.
Planta. 2002 Jul; 215(3):430-9.P

Abstract

ADP-glucose pyrophosphorylase (AGPase), a key enzyme in starch biosynthesis of higher plants, consists of a pair of regulatory large (LS) and catalytically small (SS) subunits. In plants, these subunits are coded by multiple genes resulting in the formation of tissue-specific enzyme forms, which are differentially regulated during plant growth and development. Some AGPase isoforms differ in catalytic and regulatory properties as well as intracellular location. In an effort to gain a better understanding of the role of the leaf AGPase in carbon partitioning and its effect on plant productivity, the Arabidopsis leaf AGPase containing the mature forms of the SS and LS was expressed in a heterologous expression system and characterized enzymatically. The Arabidopsis recombinant AGPase had kinetic values for 3-phosphoglyceric acid, glucose-1-phosphate and Mg(2+) similar to those of the native enzyme. As the N-terminus of the LS has been suggested to be involved in enzyme function, the length of the N-terminal region was extended or shortened. Of the five modified LSs analyzed, only the T5 form lacking six residues of the mature N-terminus was able to form detectable levels of enzyme activity, indicating that the N-terminal region is critical for enzyme function. Two up-regulatory LS mutations that allosterically activate the potato enzyme, a stem isoform, were introduced into the corresponding Arabidopsis LS sequences and co-expressed with wild-type SS. Both modified enzymes showed up-regulatory properties, indicating that these specific residue changes were also operational in the leaf isoform.

Authors+Show Affiliations

Institute of Biological Chemistry and School of Molecular Biosciences, Washington State University, Pullman, WA 99164, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12111225

Citation

Kavakli, I Halil, et al. "Generation, Characterization, and Heterologous Expression of Wild-type and Up-regulated Forms of Arabidopsis Thaliana Leaf ADP-glucose Pyrophosphorylase." Planta, vol. 215, no. 3, 2002, pp. 430-9.
Kavakli IH, Kato C, Choi SB, et al. Generation, characterization, and heterologous expression of wild-type and up-regulated forms of Arabidopsis thaliana leaf ADP-glucose pyrophosphorylase. Planta. 2002;215(3):430-9.
Kavakli, I. H., Kato, C., Choi, S. B., Kim, K. H., Salamone, P. R., Ito, H., & Okita, T. W. (2002). Generation, characterization, and heterologous expression of wild-type and up-regulated forms of Arabidopsis thaliana leaf ADP-glucose pyrophosphorylase. Planta, 215(3), 430-9.
Kavakli IH, et al. Generation, Characterization, and Heterologous Expression of Wild-type and Up-regulated Forms of Arabidopsis Thaliana Leaf ADP-glucose Pyrophosphorylase. Planta. 2002;215(3):430-9. PubMed PMID: 12111225.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Generation, characterization, and heterologous expression of wild-type and up-regulated forms of Arabidopsis thaliana leaf ADP-glucose pyrophosphorylase. AU - Kavakli,I Halil, AU - Kato,Chikako, AU - Choi,Sang-Bong, AU - Kim,Kyung-Hwan, AU - Salamone,Peter R, AU - Ito,Hiroyuki, AU - Okita,Thomas W, Y1 - 2002/04/09/ PY - 2001/09/27/received PY - 2001/11/07/accepted PY - 2002/7/12/pubmed PY - 2002/9/11/medline PY - 2002/7/12/entrez SP - 430 EP - 9 JF - Planta JO - Planta VL - 215 IS - 3 N2 - ADP-glucose pyrophosphorylase (AGPase), a key enzyme in starch biosynthesis of higher plants, consists of a pair of regulatory large (LS) and catalytically small (SS) subunits. In plants, these subunits are coded by multiple genes resulting in the formation of tissue-specific enzyme forms, which are differentially regulated during plant growth and development. Some AGPase isoforms differ in catalytic and regulatory properties as well as intracellular location. In an effort to gain a better understanding of the role of the leaf AGPase in carbon partitioning and its effect on plant productivity, the Arabidopsis leaf AGPase containing the mature forms of the SS and LS was expressed in a heterologous expression system and characterized enzymatically. The Arabidopsis recombinant AGPase had kinetic values for 3-phosphoglyceric acid, glucose-1-phosphate and Mg(2+) similar to those of the native enzyme. As the N-terminus of the LS has been suggested to be involved in enzyme function, the length of the N-terminal region was extended or shortened. Of the five modified LSs analyzed, only the T5 form lacking six residues of the mature N-terminus was able to form detectable levels of enzyme activity, indicating that the N-terminal region is critical for enzyme function. Two up-regulatory LS mutations that allosterically activate the potato enzyme, a stem isoform, were introduced into the corresponding Arabidopsis LS sequences and co-expressed with wild-type SS. Both modified enzymes showed up-regulatory properties, indicating that these specific residue changes were also operational in the leaf isoform. SN - 0032-0935 UR - https://www.unboundmedicine.com/medline/citation/12111225/Generation_characterization_and_heterologous_expression_of_wild_type_and_up_regulated_forms_of_Arabidopsis_thaliana_leaf_ADP_glucose_pyrophosphorylase_ DB - PRIME DP - Unbound Medicine ER -