Title
Detection of GTP-binding proteins in barley aleurone protoplasts
Author
Wang, M.
Sedee, N.J.A.
Heidekamp, F.
Snaar-Jagalska, B.E.
Publication year
1993
Abstract
We report the existence of several families of GTP-binding proteins in barley aleurone protoplasts. Partial purified plasma membrane proteins were separated by SDS-PAGE, transferred to a nitrocellulose filter and incubated with either antisera raised against a highly conserved animal G protein alpha subunit peptide/or Ras protein, or with [alpha-32P]GTP. Two sets of proteins of M(r) = 32-36 kDa and 22-24 kDa were strongly recognized by the antisera. Binding of [alpha-32P]GTP was detected on Western blots with proteins of M(r) = 22-24 kDa and 16 kDa. Binding was inhibited by 10-7-10-6 M GTPgammaS, GTP or GDP; binding was not affected by 10-6-10-5 M ATPgammaS or ADP. The kinetics, specificity and the effects of phytohormones in a[35S]GTPgammaS binding assay were also studied in isolated plasma membranes of barley aleurone protoplasts. Chemicals/CAS: GTP-Binding Proteins, EC 3.6.1.-; Plant Growth Regulators
Subject
Barley aleurone
GTP-binding protein
Phytohormone
Guanine nucleotide binding protein
Phytohormone
Barley
Nonhuman
Protein purification
Protoplast
Binding Sites
Blotting, Western
Cell Membrane
Electrophoresis, Polyacrylamide Gel
GTP-Binding Proteins
Hordeum
Plant Growth Regulators
Protoplasts
Signal Transduction
Support, Non-U.S. Gov't
Temperature
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DOI
https://doi.org/10.1016/0014-5793(93)80230-r
TNO identifier
36713
Source
FEBS Letters, 329 (329), 245-248
Document type
article