Mapping of Chlamydia trachomatis proteins by immobiline-polyacrylamide two-dimensional electrophoresis: spot identification by N-terminal sequencing and immunoblotting.
Proteins from purified elementary bodies of Chlamydia trachomatis were separated by two-dimensional gel electrophoresis on nonlinear wide-range immobilized pH gradients in the first dimension and polyacrylamide gradient gels in the second dimension. The maps obtained with this system are highly reproducible and resolve ca. 600 spots. By using immunoblot analysis with specific antibodies and/or N-terminal amino acid sequencing, we established the map positions of a number of described chlamydial proteins, such as the major outer membrane protein (MOMP) the 60 kDa cystein-rich outer membrane protein (OMP2), the DnaK-like, GroEL-like, and macrophage infectivity potentiator (MIP)-like proteins, the plasmid-encoded pgp3 protein, two ribosomal proteins (S1 and L7/L12), and the protein-elongation factor EF-Tu. Other proteins, for which gene assignment was not possible, have been identified by three parameters (Mr, pI and N-terminal sequence). This work provides a preliminary basis for a future and progressive compilation of a genome-linked database of chlamydial proteins.
Department of Molecular Biology, Siena University, Italy.
L
L
Bini
M
M
Sanchez-Campillo
A
A
Santucci
B
B
Magi
B
B
Marzocchi
M
M
Comanducci
G
G
Christiansen
S
S
Birkelund
R
R
Cevenini
E
E
Vretou
G
G
Ratti
V
V
Pallini
D000178:Acrylamides; D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis; D001426:Bacterial Proteins / Q000737:chemistry*; D002692:Chlamydia trachomatis; D015180:Electrophoresis, Gel, Two-Dimensional; D016680:Genome, Bacterial; D015151:Immunoblotting; D008832:Microchemistry; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods*; D016622:Silver Staining
D000178:Acrylamides; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; C059413:Immobiline