Truncation of recombinant vimentin by ompT. Identification of a short motif in the head domain necessary for assembly of type III intermediate filament proteins.
Recombinant vimentin expressed in E. coli JM 101 cells is cleaved after cell lysis between arginines 11 and 12. The truncated vimentin is assembly incompetent. Expression of the same cDNA construct in BL21 cells, which lack the protease ompT, provides intact and polymerization-competent vimentin. The ompT cleavage site is contained in a short sequence motif (YRRMF) shared by the head domains of type III and IV intermediate filament (IF) proteins. We propose that a related motif present in the N-terminal 32 residues of lambda CII accounts for the known IF formation of a fusion protein formed with a truncated GFAP.
Hatzfeld
M
M
Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Goettingen, Germany.
Dodemont
H
H
Plessmann
U
U
Weber
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005904:Glial Fibrillary Acidic Protein / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000378:metabolism; D014746:Vimentin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; D011994:Recombinant Proteins; D014746:Vimentin; D004247:DNA