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Lateral diffusion of the secretory component (SC) in the basolateral membrane of the human colon carcinoma cell line HT29 assessed with fluorescence recovery after photobleaching
Lateral diffusion of the secretory component (SC) in the basolateral membrane of the human colon carcinoma cell line HT29 assessed with fluorescence recovery after photobleaching
- Source :
- Journal of cellular physiology. 137(3)
- Publication Year :
- 1988
-
Abstract
- The lateral diffusion of the secretory component (SC), acting as a receptor for dimeric IgA in the basolateral side of intestinal epithelial cells, was studied in the human colonic carcinoma cell line HT29. The HT29 cells were grown in Dulbecco's modified Eagle's medium in which galactose had been substituted for glucose to promote development of small intestine-like cells, with a distinct separation of the basolateral side from the apical surface. The SC was stained with rhodamine-labeled polyclonal anti-human SC rabbit antibodies (Ig) or Fab fragments, and the lateral mobility was assessed with the fluorescence recovery after photobleaching technique. The average lateral diffusion was consistent with a diffusion constant of 7.7 +/- 2.0 (mean value +/- SD; n = 29) and 7.1 +/- 2.3 (n = 30) x 10(-10) cm2s-1 for Ig-and Fab-labeled receptors, respectively, which is slower than lipid diffusion but is similar to that found for other membrane receptors. The corresponding values for the fraction of mobile receptors were 66 +/- 13% and 71 +/- 12%, respectively. Cells were labeled from the top of the culture plate, and cells adjacent to a mechanically made rift or a natural opening in the cell monolayer were labeled more strongly, confirming the microscope-based impression that the basolateral surface primarily harboured the SC receptor.
- Subjects :
- Physiology
Secretory component
Clinical Biochemistry
Fluorescence recovery after photobleaching
Cell Biology
Receptors, Fc
Biology
Fluorescence
Secretory Component
Diffusion
HT29 Cells
Biochemistry
Cell surface receptor
Cell culture
Polyclonal antibodies
Colonic Neoplasms
biology.protein
Biophysics
Tumor Cells, Cultured
Humans
Intestinal Mucosa
Receptors, Immunologic
Receptor
Immunoglobulin Fragments
Epithelial polarity
Subjects
Details
- ISSN :
- 00219541
- Volume :
- 137
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Journal of cellular physiology
- Accession number :
- edsair.doi.dedup.....13446970de96e88fadd6a5816b5dedc7