Biochemistry, Biophysics, and Structural Biology Commons^™

Kinetic Properties Of Bifunctional 6-Phosphofructo-2-Kinase/Fructose-2,6-Bisphosphatase From Spinach Leaves, Jennifer E. Markham, Nicholas J. Kruger

Department of Biochemistry: Faculty Publications

A cDNA encoding 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase was isolated from a Spinacia oleracea leaf library and used to ex­press a recombinant enzyme in Escherichia coli and Spodoptera frugiperda cells. The insoluble protein expressed in E. coli was purified and used to raise antibodies. Western blot analysis of a protein extract from spinach leaf showed a single band of 90.8 kDa. Soluble protein was purified to homogeneity from S. frugiperda cells infected with recombinant baculovirus harboring the isolated cDNA. The soluble protein had a molecular mass of 320 kDa, estimated by gel filtration chromatography, and a subunit size of 90.8 kDa. The purified pro­tein …