A (His)6‐tagged recombinant barley (Hordeum vulgare L.) endosperm ADP‐glucose pyrophosphorylase expressed in the baculovirus‐insect cell system is insensitive to allosteric regulation by 3‐phosphoglycerate and inorganic phosphate

FEBS Letters

Doan, Danny N.P¹  Rudi, Heidi¹  Olsen, Odd-Arne¹ 

[1] Plant Molecular Biology Laboratory, Department of Biotechnological Sciences, Agricultural University of Norway, P.O. Box 5051, N-1432 Ås, Norway

关键词: ADP-glucose pyrophosphorylase;    Expression;    Baculovirus;    Insect cell;    Purification;   

DOI  :  10.1016/S0014-5793(97)01448-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

ADP-glucose pyrophosphorylase from photosynthetic tissue is allosterically regulated by 3-phosphoglycerate and inorganic phosphate. In contrast, data from our laboratory indicated that the major AGPase from barley seeds is insensitive to these regulators. Verification of this conclusion has, however, been hindered by the proteolytic degradation of the enzyme from seeds. This report characterizes the barley seed AGPase expressed in the baculovirus-insect cell system, confirming that lack of allosteric regulation by 3-PGA/P_i is an intrinsic property of the enzyme. Purification of the enzyme was by Ni²⁺-NTA agarose chromatography using a (His)₆ tag attached to the N-terminus of the small AGPase subunit.

【 授权许可】

Unknown