title: Comparative proteome analysis of Chlamydia trachomatis serovar A, D and L2.
abstract: Chlamydia trachomatis represents a group of human pathogenic obligate intracellular and gram-negative bacteria. The genome of C. trachomatis D comprises 894 open reading frames (ORFs). In this study the global expression of genes in C. trachomatis A, D and L2, which are responsible for different chlamydial diseases, was investigated using a proteomics approach. Based on silver stained two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), gels with purified elementary bodies (EB) and auto-radiography of gels with 35S-labeled C. trachomatis proteins up to 700 protein spots were detectable within the range of the immobilized pH gradient (IPG) system used. Using mass spectrometry and N-terminal sequencing followed by database searching we identified 250 C. trachomatis proteins from purified EB of which 144 were derived from different genes representing 16% of the ORFs predicted from the C. trachomatis D genome and the 7.5 kb C. trachomatis plasmid. Important findings include identification of proteins from the type III secretion apparatus, enzymes from the central metabolism and confirmation of expression of 25 hypothetical ORFs and five polymorphic membrane proteins. Comparison of serovars generated novel data on genetic variability as indicated by electrophoretic variation and potentially important examples of serovar specific differences in protein abundance. The availability of the complete genome made it feasible to map and to identify proteins of C. trachomatis on a large scale and the integration of our data in a 2-D PAGE database will create a basis for post genomic research, important for the understanding of chlamydial development and pathogenesis.
authors.affiliation: Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus, Denmark. shaw@medmicro.aau.dk
authors.forename: Allan C
authors.initials: AC
authors.lastname: Shaw
authors.forename: Kris
authors.initials: K
authors.lastname: Gevaert
authors.forename: Hans
authors.initials: H
authors.lastname: Demol
authors.forename: Bart
authors.initials: B
authors.lastname: Hoorelbeke
authors.forename: Joël
authors.initials: J
authors.lastname: Vandekerckhove
authors.forename: Martin R
authors.initials: MR
authors.lastname: Larsen
authors.forename: Peter
authors.initials: P
authors.lastname: Roepstorff
authors.forename: Arne
authors.initials: A
authors.lastname: Holm
authors.forename: Gunna
authors.initials: G
authors.lastname: Christiansen
authors.forename: Svend
authors.initials: S
authors.lastname: Birkelund
mesh_terms: D001345:Autoradiography; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D002692:Chlamydia trachomatis / Q000737:chemistry* / Q000145:classification / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression; D005798:Genes, Bacterial; D014644:Genetic Variation; D006367:HeLa Cells; D006801:Humans; D020543:Proteome / Q000032:analysis* / Q000235:genetics; D012703:Serotyping; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001426:Bacterial Proteins; D020543:Proteome