Purification and characterisation of an antifreeze protein from Forsythia suspensa (L.).
Recrystallisation inhibition (RI) activity has been isolated from cold-acclimated Forsythia suspensa bark and leaves, which is stable when boiled, and not sensitive to reducing agents. The antifreeze activity has been purified to a single 20 kDa protein, using anion exchange, hydroxyapatite chromatography, and gel filtration. The protein is abundant in forsythia bark with 0.5microg pure protein obtained from 35 g bark. RI activity is seen with as little as 6 microg ml(-1) protein. Sequence homology was seen with dehydrins, and forsythia AFP contains the Y-segment, a conserved region found in many dehydrins.
Simpson
Deborah J
DJ
Cardiff School of Biosciences, Main Building, Cardiff University, Museum Avenue, Cardiff CF10 3TL, UK. simpsondj@cardiff.ac.uk
Smallwood
Maggie
M
Twigg
Sarah
S
Doucet
Charlotte J
CJ
Ross
Joe
J
Bowles
Dianna J
DJ
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D021301:Antifreeze Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods; D015925:Cryopreservation / Q000379:methods; D031657:Forsythia / Q000737:chemistry*; D008969:Molecular Sequence Data; D024301:Plant Bark / Q000737:chemistry; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification; D013997:Time Factors
D000596:Amino Acids; D021301:Antifreeze Proteins; D010940:Plant Proteins; C070556:dehydrin proteins, plant