Analysis of post-translational modification of mycobacterial proteins using a cassette expression system.
A recombinant expression system was developed to analyse sequence determinants involved in O-glycosylation of proteins in mycobacteria. By expressing peptide sequences corresponding to known glycosylation sites within a chimeric lipoprotein construct, amino acids flanking modified threonine residues were found to have an important influence on glycosylation. The expression system was used to screen mycobacterial sequences selected using a neural network (NetOglyc) trained on eukaryotic O-glycoproteins. Evidence of glycosylation was obtained for eight of 11 proteins tested. The results suggest that sites involved in O-glycosylation of mycobacterial and eukaryotic proteins share similar structural features.
Herrmann
J L
JL
Department of Infectious Diseases and Microbiology, Imperial College School of Medicine, St. Mary's Campus, Norfolk Place, W2 1PG, London, UK. jlh@chu-stlouis.fr
Delahay
R
R
Gallagher
A
A
Robertson
B
B
Young
D
D
D000469:Alkaline Phosphatase / Q000235:genetics; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D003208:Concanavalin A / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics; D006031:Glycosylation; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics* / Q000378:metabolism; D016571:Neural Networks, Computer; D011499:Protein Processing, Post-Translational; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D004274:DNA, Recombinant; D008074:Lipoproteins; D003208:Concanavalin A; D000469:Alkaline Phosphatase