Recombinant tobacco mosaic virus movement protein is an RNA-binding, alpha-helical membrane protein.
The 30-kDa movement protein (MP) is essential for cell-cell spread of tobacco mosaic virus in planta. To explore the structural properties of MP, the full-length recombinant MP gene was expressed in Escherichia coli, and one-step purification from solubilized inclusion bodies was accomplished by using anion exchange chromatography. Soluble MP was maintained at >4 mg/ml without aggregation and displayed approximately 70% alpha-helical conformation in the presence of urea and SDS. A trypsin-resistant core domain of the MP had tightly folded tertiary structure, whereas 18 aa at the C terminus of the monomer were rapidly removed by trypsin. Two hydrophobic regions within the core were highly resistant to proteolysis. Based on results of CD spectroscopy, trypsin treatment, and MS, we propose a topological model in which MP has two putative alpha-helical transmembrane domains and a protease-sensitive carboxyl terminus.
Brill
L M
LM
Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, Mail Drop MB 28, La Jolla, CA 92037, USA.
Nunn
R S
RS
Kahn
T W
TW
Yeager
M
M
Beachy
R N
RN
D000595:Amino Acid Sequence; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D053830:Plant Viral Movement Proteins; D017510:Protein Folding; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014027:Tobacco Mosaic Virus / Q000235:genetics* / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D008565:Membrane Proteins; D053830:Plant Viral Movement Proteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins