Physicochemical and kinetic properties of unique isozymes of UDP-Glc pyrophosphorylase that are associated with resistance to sweetening in cold-stored potato tubers.
Isozymes of UGPase with unique catalytic properties were purified from the cold-induced-sweetening (CIS) resistant cultivar Snowden (Solanum tuberosum). Two distinct peaks of UGPase activity were obtained when protein extracts were subjected to anion-exchange chromatography on DEAE-Sephacel. Polypeptides in the first eluted fraction (A-I) were ionically similar to the UGPase isozyme UGP3 previously purified and characterized from the cold-sweetening sensitive cultivar Norchip (Sowokinos et al. 1993, Plant Physiol 101: 1073-1080). Seventy-two percent of the total endogenous UGPase activity in Snowden (cv.) tubers, however, was found in a more basic protein fraction (A-II) that is not found in the Norchip cultivar. This study reports on the physicochemical and kinetic properties of these new polypeptides that demonstrate UGPase activity. The reaction in the direction of UDP-Glc synthesis was specific for the substrates Glc-1-P and UTP and there was an absolute requirement for Mg2+ ions. The catalytic properties of UGP5 were markedly different from UGPase isozymes previously described in terms of (1) affinity for the substrate Glc-1-P, (2) pH optimum, (3) maximum reaction velocity and (4) sensitivity to product inhibition with UDP-Glc. Chi-square analysis of fifty-four genetically diverse potato lines revealed that resistance to CIS was highly correlated with the presence of the A-II isozymes of UGPase. The kinetic properties of these unique forms of UGPase may underlie, in part, a tuber's ability to resist sweetening in the cold.
Gupta
Sanjay Kumar
SK
University of Minnesota, Department of Horticultural Science, St. Paul, MN 55108, USA.
Sowokinos
Joseph Robin
JR
D000064:Acclimatization / Q000502:physiology; D002241:Carbohydrates / Q000096:biosynthesis*; D003080:Cold Temperature; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D018547:Plant Stems / Q000201:enzymology*; D011198:Solanum tuberosum / Q000201:enzymology* / Q000502:physiology; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000302:isolation & purification / Q000378:metabolism*
D002241:Carbohydrates; D007527:Isoenzymes; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase