Amino acid sequence of spinach chloroplast fructose-1,6-bisphosphatase.
The amino acid sequence of the spinach chloroplast fructose-1,6-bisphosphatase (FBPase) subunit has been determined. Placement of the 358 residues in the polypeptide chain was based on automated Edman degradation of the intact protein and of peptides obtained by enzymatic or chemical cleavage. The sequence of spinach chloroplast FBPase shows clear homology (ca. 40%) to gluconeogenic (mammalian, yeast, and Escherichia coli) fructose-1,6-bisphosphatases and 80% homology with the wheat chloroplast enzyme. The two chloroplast enzymes show near the middle of the structure a unique sequence insert probably involved in light-dependent regulation of the chloroplast FBPase enzyme activity. This sequence insert contains two cysteines separated by only 4 amino acid residues, a characteristic feature of some enzymes containing redox-active cysteines. The recent X-ray crystallographic resolution of pig kidney FBPase (H. Ke, C. M. Thorpe, B. A. Seaton, F. Marcus, and W. N. Lipscomb, 1989, Proc. Natl. Acad. Sci. USA 86, 1475-1479) has allowed the discussion of the amino acid sequence of spinach chloroplast FBPase in structural terms. It is to be noted that most of pig kidney FBPase residues shown to be either at (or close to) the sugar bisphosphate binding site or located at the negatively charged metal binding pocket are conserved in the chloroplast enzyme. The unique chloroplast FBPase insert presumably involved in light-dependent activation of the enzyme via a thioredoxin-linked mechanism can be accommodated in the surface of the FBPase molecule.
Marcus
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Department of Biological Chemistry and Structure, University of Health Sciences, Chicago Medical School.
Harrsch
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PB
D000595:Amino Acid Sequence; D000818:Animals; D002736:Chloroplasts / Q000201:enzymology*; D006597:Fructose-Bisphosphatase / Q000032:analysis*; D007668:Kidney / Q000201:enzymology; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology*; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013552:Swine
D006597:Fructose-Bisphosphatase