Molecular basis for thermoprotection in Bemisia: structural differences between whitefly ketose reductase and other medium-chain dehydrogenases/reductases.
The silverleaf whitefly (Bemisia argentifolii, Bellows and Perring) accumulates sorbitol as a thermoprotectant in response to elevated temperature. Sorbitol synthesis in this insect is catalyzed by an unconventional ketose reductase (KR) that uses NADPH to reduce fructose. A cDNA encoding the NADPH-KR from adult B. argentifolii was cloned and sequenced to determine the primary structure of this enzyme. The cDNA encoded a protein of 352 amino acids with a calculated molecular mass of 38.2 kDa. The deduced amino acid sequence of the cDNA shared 60% identity with sheep NAD(+)-dependent sorbitol dehydrogenase (SDH). Residues in SDH involved in substrate binding were conserved in the whitefly NADPH-KR. An important structural difference between the whitefly NADPH-KR and NAD(+)-SDHs occurred in the nucleotide-binding site. The Asp residue that coordinates the adenosyl ribose hydroxyls in NAD(+)-dependent dehydrogenases (including NAD(+)-SDH), was replaced by an Ala in the whitefly NADPH-KR. The whitefly NADPH-KR also contained two neutral to Arg substitutions within four residues of the Asp to Ala substitution. Molecular modeling indicated that addition of the Arg residues and loss of the Asp decreased the electric potential of the adenosine ribose-binding pocket, creating an environment favorable for NADPH-binding. Because of the ability to use NADPH, the whitefly NADPH-KR synthesizes sorbitol under physiological conditions, unlike NAD(+)-SDHs, which function in sorbitol catabolism.
Wolfe
G R
GR
Western Cotton Research Laboratory, US Department of Agriculture, Agricultural Research Service, Phoenix, AZ 85040, USA.
Smith
C A
CA
Hendrix
D L
DL
Salvucci
M E
ME
D019747:Aldose-Ketose Isomerases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine; D001833:Body Temperature Regulation / Q000502:physiology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D006430:Hemiptera / Q000502:physiology*; D007064:L-Iditol 2-Dehydrogenase / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009249:NADP; D011485:Protein Binding; D017421:Sequence Analysis; D013012:Sorbitol / Q000378:metabolism*; D013696:Temperature
D018076:DNA, Complementary; D013012:Sorbitol; D009249:NADP; D001120:Arginine; D007064:L-Iditol 2-Dehydrogenase; D019747:Aldose-Ketose Isomerases