Purification of 1-aminocyclopropane-1-carboxylate synthase from apple fruits using s-adenosyl (3,4 sup 14 C)-methionine (SAM) as a probe

Journal Article · Sat Apr 01 04:00:00 EST 1989 · Plant Physiology, Supplement; (USA)

OSTI ID:6611042

Yip, Wingkip; Dong, Jianguo; Yang, Shang Fa ^[1]

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1. Univ. of California, Davis (USA)

Tomato ACC synthase is inactivated by its substrate SAM, with the moiety of aminobutyrate being covalently linked to ACC synthase during the catalytic reactions. A partial purified ACC synthase (the catalytic activity 100 {mu}mol/h{center dot}mg protein) from pellets of apple extract was incubated with (3,4{sup 14}C) SAM. Only one radioactive peak was revealed in a C-4 reverse phase HPLC and one radioactive band on SDS-PAGE with an M.W. of 48 kDa. Apple ACC synthase in native form is resistant to V8, {alpha}-chromtrypsin and carboxylpeptidase A digestion, but effectively inactivated by trypsin and ficin, as demonstrated by both the activity assay and SAM labeling. The radioactive protein cut from the SDS-PAGE was injected to three mice, two of the mice showed responses to the protein in western blot analysis. The antibodies from mice is currently under characterization.

• OSTI ID: 6611042
• Journal Information: Plant Physiology, Supplement; (USA), Journal Name: Plant Physiology, Supplement; (USA) Vol. 89:4; ISSN PPYSA; ISSN 0079-2241
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
APPLES
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CHROMATOGRAPHY
DRUGS
ELECTROPHORESIS
ENZYMES
FOOD
FRACTIONATION
FRUITS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIGASES
LIPOTROPIC FACTORS
LIQUID COLUMN CHROMATOGRAPHY
METHIONINE
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
SEPARATION PROCESSES
TRACER TECHNIQUES