TY - JOUR
T1 - Molecular cloning, sequence analysis, and expression of a human liver cDNA coding for fructose-6-P,2-kinase:Fructose-2,6-bisphosphatase
AU - Algaier, Joseph
AU - Uyeda, Kosaku
N1 - Funding Information:
ACKNOWLEDGMENT: This work was supported by the Veterans Administration and by United States Public Health Service Grant No. AM16194. We thank Dr. William J. Rutter (Univ. of California-San Francisco) for providing human liver xgt11 cDNA library and Dr. Stan Tabor for pT7-7 RNA polymerase/promoter plasmid. We also thank Barbara Beard for her skillful technical assistance.
PY - 1988/5/31
Y1 - 1988/5/31
N2 - A cDNA coding for 378 amino acids from the C-terminus of the human liver bifunctional enzyme, Fructose-6-phosphate,2-kinase:Fructose-2,6-bisphosphatase was isolated, sequenced, and expressed in E. coli K38. The expressed protein, identified by specific immunoassay, showed Fru 2,6-bisphosphatase activity but no Fru 6-P,2-kinase activity, demonstrating directly that the Fru 2,6-bisphosphatase activity resides in the C-terminal region. The Km for Fru 2,6-P2 was 4.3 μM. Fru 6-P was a noncompetitive inhibitor (Ki = 2.9 μM), and formed a phosphorylated intermediate when incubated with Fru 2,6[2-32P]P2. The subunit Mr of the enzyme was 36,600, and the active enzyme showed Mr=37,000 by gel filtration.
AB - A cDNA coding for 378 amino acids from the C-terminus of the human liver bifunctional enzyme, Fructose-6-phosphate,2-kinase:Fructose-2,6-bisphosphatase was isolated, sequenced, and expressed in E. coli K38. The expressed protein, identified by specific immunoassay, showed Fru 2,6-bisphosphatase activity but no Fru 6-P,2-kinase activity, demonstrating directly that the Fru 2,6-bisphosphatase activity resides in the C-terminal region. The Km for Fru 2,6-P2 was 4.3 μM. Fru 6-P was a noncompetitive inhibitor (Ki = 2.9 μM), and formed a phosphorylated intermediate when incubated with Fru 2,6[2-32P]P2. The subunit Mr of the enzyme was 36,600, and the active enzyme showed Mr=37,000 by gel filtration.
UR - http://www.scopus.com/inward/record.url?scp=0023947330&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0023947330&partnerID=8YFLogxK
U2 - 10.1016/S0006-291X(88)81226-9
DO - 10.1016/S0006-291X(88)81226-9
M3 - Article
C2 - 2837207
AN - SCOPUS:0023947330
SN - 0006-291X
VL - 153
SP - 328
EP - 333
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -