Cold stability of pyruvate, orthophosphate dikinase of Flaveria brownii.
The nucleotide sequences of the complementary DNA of pyruvate, Pi dikinase (PPDK) from Flaveria bidentis, a C4 plant which possesses a cold-sensitive form of PPDK, and Flaveria brownii, a 'C4-like' plant which possesses a cold-tolerant form of PPDK, were determined. PPDK was isolated from the leaves of both Flaveria species and purified and the N-terminal amino acid sequences characterised. Together with a maize PPDK cDNA, cDNA inserts which code for the mature form of PPDK of F. bidentis and of F. brownii were expressed in bacteria and the cold sensitivity of the expressed PPDK studied. The cold sensitivity of the PPDK expressed in bacteria mimics the cold sensitivity of PPDK found in vivo in all three plant species. This study indicates that the cold sensitivity of plant PPDK is controlled by the primary structure of the enzyme.
Usami
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S
Plant Breeding and Genetics Research Laboratory, Japan Tobacco Inc., Shizuoka.
Ohta
S
S
Komari
T
T
Burnell
J N
JN
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003080:Cold Temperature; D017931:DNA Primers; D018076:DNA, Complementary / Q000737:chemistry; D004795:Enzyme Stability; D015870:Gene Expression; D007700:Kinetics; D008969:Molecular Sequence Data; D018515:Plant Leaves; D010944:Plants / Q000201:enzymology* / Q000235:genetics; D011772:Pyruvate, Orthophosphate Dikinase / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; D011772:Pyruvate, Orthophosphate Dikinase