TY - JOUR
T1 - Specific detection of Salmonella typhimurium proteins synthesized intracellularly
AU - Burns-Keliher, Lisa L.
AU - Portteus, Andrew
AU - Curtiss, Roy
PY - 1997/6
Y1 - 1997/6
N2 - Studies of the proteins Salmonella typhimurium synthesizes under conditions designed to more closely approximate the in vivo environment, i.e., in cell and tissue culture, are not easily interpreted because they have involved chemical inhibition of host cell protein synthesis during infection. The method which we have developed allows specific labeling of bacterial proteins without interfering with host cell metabolic activities by using a labeled lysine precursor which mammalian cells cannot utilize. We have resolved the labeled proteins using two-dimensional electrophoresis and autofluorography. We were able to detect 57 proteins synthesized by S. typhimurium during growth within a human intestinal epithelial cell line. Of the 57 proteins detected, 34 appear to be unique to the intracellular environment, i.e., they are not seen during growth of the bacteria in tissue culture medium alone. Current (and future) efforts are directed at organizing the 34 proteins into known stress response groups, determining the cellular locations of the proteins (outer or inner membrane, etc.), and comparing the pattern of proteins synthesized within an intestinal epithelial cell to the pattern synthesized during growth within other tissues.
AB - Studies of the proteins Salmonella typhimurium synthesizes under conditions designed to more closely approximate the in vivo environment, i.e., in cell and tissue culture, are not easily interpreted because they have involved chemical inhibition of host cell protein synthesis during infection. The method which we have developed allows specific labeling of bacterial proteins without interfering with host cell metabolic activities by using a labeled lysine precursor which mammalian cells cannot utilize. We have resolved the labeled proteins using two-dimensional electrophoresis and autofluorography. We were able to detect 57 proteins synthesized by S. typhimurium during growth within a human intestinal epithelial cell line. Of the 57 proteins detected, 34 appear to be unique to the intracellular environment, i.e., they are not seen during growth of the bacteria in tissue culture medium alone. Current (and future) efforts are directed at organizing the 34 proteins into known stress response groups, determining the cellular locations of the proteins (outer or inner membrane, etc.), and comparing the pattern of proteins synthesized within an intestinal epithelial cell to the pattern synthesized during growth within other tissues.
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U2 - 10.1128/jb.179.11.3604-3612.1997
DO - 10.1128/jb.179.11.3604-3612.1997
M3 - Article
C2 - 9171406
AN - SCOPUS:0030990845
SN - 0021-9193
VL - 179
SP - 3604
EP - 3612
JO - Journal of bacteriology
JF - Journal of bacteriology
IS - 11
ER -