Expression of barley ADP-glucose pyrophosphorylase in Escherichia coli: processing and regulatory considerations. Luo, C. & Kleczkowski, L. A Phytochemistry, 50(2):209–214, January, 1999.
Expression of barley ADP-glucose pyrophosphorylase in Escherichia coli: processing and regulatory considerations [link]Paper  doi  abstract   bibtex   
Full length cDNAs for barley ADP-glucose pyrophosphorylase (AGPase) coding for the large subunits of the endosperm and leaf homologues of the enzyme (AGPase-S1 and -S2, respectively) and for the small subunit protein from endosperm (AGPase-B1), have been expressed in Escherichia coli. The cDNAs for AGPase-S1 and -S2 required different induction conditions for their maximal expression and they encoded immunologically distinct proteins. The AGPase-S1 that was produced by E. coli had the same Mr (58 kDa) as the corresponding protein in barley crude endosperm extracts, whereas the bacteria-produced AGPase-S2 (55 kDa) was larger than its counterpart from barley leaf preparations (53 kDa). An enzymatically active AGPase expressed in E. coli from a double construct containing cDNAs for AGPase-S1 and -B1 subunits was insensitive to the activation by 3-phosphoglycerate and to inhibition by inorganic phosphate, similarly to the enzyme in barley endosperm. Neither AGPase-S1 nor -B1 were active when expressed alone in the bacteria. The data are discussed with respect to possible mechanisms of intracellular targeting of immature AGPase-S proteins in barley tissues and regarding previous data on effector regulation of the barley enzyme.
@article{luo_expression_1999,
	title = {Expression of barley {ADP}-glucose pyrophosphorylase in {Escherichia} coli: processing and regulatory considerations},
	volume = {50},
	issn = {0031-9422},
	shorttitle = {Expression of barley {ADP}-glucose pyrophosphorylase in {Escherichia} coli},
	url = {https://www.sciencedirect.com/science/article/pii/S0031942298004725},
	doi = {10/dck4r3},
	abstract = {Full length cDNAs for barley ADP-glucose pyrophosphorylase (AGPase) coding for the large subunits of the endosperm and leaf homologues of the enzyme (AGPase-S1 and -S2, respectively) and for the small subunit protein from endosperm (AGPase-B1), have been expressed in Escherichia coli. The cDNAs for AGPase-S1 and -S2 required different induction conditions for their maximal expression and they encoded immunologically distinct proteins. The AGPase-S1 that was produced by E. coli had the same Mr (58 kDa) as the corresponding protein in barley crude endosperm extracts, whereas the bacteria-produced AGPase-S2 (55 kDa) was larger than its counterpart from barley leaf preparations (53 kDa). An enzymatically active AGPase expressed in E. coli from a double construct containing cDNAs for AGPase-S1 and -B1 subunits was insensitive to the activation by 3-phosphoglycerate and to inhibition by inorganic phosphate, similarly to the enzyme in barley endosperm. Neither AGPase-S1 nor -B1 were active when expressed alone in the bacteria. The data are discussed with respect to possible mechanisms of intracellular targeting of immature AGPase-S proteins in barley tissues and regarding previous data on effector regulation of the barley enzyme.},
	language = {en},
	number = {2},
	urldate = {2021-11-08},
	journal = {Phytochemistry},
	author = {Luo, Cheng and Kleczkowski, Leszek A},
	month = jan,
	year = {1999},
	keywords = {Barley, Heterologous expression, Starch synthesis, Transit peptide},
	pages = {209--214},
}

Downloads: 0