Eimeria necatrix: induced proteolytic sensitivity of infected chick crypt cells.
While devising a protocol for the isolation of chick crypt cells infected with Eimeria necatrix, it was observed that infected cells were readily lysed by 0.25% trypsin. Time-course studies at 17 C with 5.5 x 10(5) cells at 96 hr postinfection revealed that 0.001% trypsin effectively lysed greater than 90% of infected cells within 10 min. Uninfected crypt cells were not lysed under these conditions. To determine the site of action of trypsin, the plasma membrane proteins from trypsin-treated and untreated infected cells were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. While the major proteins were unaffected by the trypsin treatment, some minor changes were noted: (1) three components (congruent to 60, congruent to 52, and congruent to 20 KDa) were trypsin sensitive and (2) a new band (congruent to 42 KDa) appeared in the membrane of trypsin-treated infected cells. Previously, it was found that the plasma membrane of infected cells, in contrast to uninfected cells, accumulated gel-phase lipid (J. E. Thompson, M. A. Fernando, and J. Pasternak, Biochimica et Biophysica Acta 555, 472-487, 1979). Here, it was examined whether trypsin would perturb the physical state of the plasma membrane of infected cells. Both X-ray diffraction patterns and transition temperature studies revealed no difference between membranes from untreated and trypsin-treated infected cells. Thus, "trypsin sensitivity" may be a secondary phenomenon that is due primarily to the cellular leakiness that accompanies the accumulation of gel-phase lipid in the plasma membrane of infected cells. The uptake of trypsin may stimulate the release of catabolic enzymes that, consequently, lyse an infected cell.
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004539:Eimeria / Q000502:physiology*; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects / Q000469:parasitology*; D008560:Membrane Fluidity / Q000187:drug effects; D008563:Membrane Lipids; D008565:Membrane Proteins / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology*
D008563:Membrane Lipids; D008565:Membrane Proteins; D014357:Trypsin