Abstract
We have examined the interactions between various radiolabeled membrane fractions obtained from an enterotoxigenic Escherichia coli strain and brush borders isolated from porcine intestinal epithelial cells. Outer membrane fragments containing the K88 attachment factor bound tightly to brush borders, whereas cytoplasmic membrane vesicles did not. Three different types of outer membrane preparations were tested: (i) cellular outer membranes isolated from lysozyme spheroplasts, (ii) medium vesicles or outer membrane fragments released into the medium during growth, and (iii) periplasmic vesicles, or outer membrane fragments which were released from the cells during spheroplast formation and were therefore isolated in the periplasmic fraction. Of these fractions, which were heterogeneous, it was always the outer membrane subfraction which bound tightly to brush borders. This binding, which was K88 dependent, may have some physiological significance in view of the association between outer membrane fragments and enterotoxin. Thus, released outer membrane fragments equipped with attachment factors may function as enterotoxin carriers which increase the efficiency with which enterotoxin can be delivered to intestinal epithelial cells.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Clements J. D., Finkelstein R. A. Isolation and characterization of homogeneous heat-labile enterotoxins with high specific activity from Escherichia coli cultures. Infect Immun. 1979 Jun;24(3):760–769. doi: 10.1128/iai.24.3.760-769.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Leij L., Witholt B. Structural heterogeneity of the cytoplasmic and outer membranes of Escherichia coli. Biochim Biophys Acta. 1977 Nov 15;471(1):92–104. doi: 10.1016/0005-2736(77)90396-0. [DOI] [PubMed] [Google Scholar]
- DiRienzo J. M., Nakamura K., Inouye M. The outer membrane proteins of Gram-negative bacteria: biosynthesis, assembly, and functions. Annu Rev Biochem. 1978;47:481–532. doi: 10.1146/annurev.bi.47.070178.002405. [DOI] [PubMed] [Google Scholar]
- Evans E. M., Wrigglesworth J. M., Burdett K., Pover W. F. Studies on epithelial cells isolated from guinea pig small intestine. J Cell Biol. 1971 Nov;51(21):452–464. doi: 10.1083/jcb.51.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finkelstein R. A., Vasil M. L., Jones J. R., Anderson R. A., Barnard T. Clinical cholera caused by enterotoxigenic Escherichia coli. J Clin Microbiol. 1976 Mar;3(3):382–384. doi: 10.1128/jcm.3.3.382-384.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Forstner G. G., Sabesin S. M., Isselbacher K. J. Rat intestinal microvillus membranes. Purification and biochemical characterization. Biochem J. 1968 Jan;106(2):381–390. doi: 10.1042/bj1060381. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gankema H., Wensink J., Guinée P. A., Jansen W. H., Witholt B. Some characteristics of the outer membrane material released by growing enterotoxigenic Escherichia coli. Infect Immun. 1980 Aug;29(2):704–713. doi: 10.1128/iai.29.2.704-713.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Germanier R., Fürer E., Varallyay S., Inderbitzin T. M. Preparation of a purified antigenic cholera toxoid. Infect Immun. 1976 Jun;13(6):1692–1698. doi: 10.1128/iai.13.6.1692-1698.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoekstra D., van der Laan J. W., de Leij L., Witholt B. Release of outer membrane fragments from normally growing Escherichia coli. Biochim Biophys Acta. 1976 Dec 14;455(3):889–899. doi: 10.1016/0005-2736(76)90058-4. [DOI] [PubMed] [Google Scholar]
- Jones G. W., Rutter J. M. Role of the K88 antigen in the pathogenesis of neonatal diarrhea caused by Escherichia coli in piglets. Infect Immun. 1972 Dec;6(6):918–927. doi: 10.1128/iai.6.6.918-927.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kessler M., Acuto O., Storelli C., Murer H., Müller M., Semenza G. A modified procedure for the rapid preparation of efficiently transporting vesicles from small intestinal brush border membranes. Their use in investigating some properties of D-glucose and choline transport systems. Biochim Biophys Acta. 1978 Jan 4;506(1):136–154. doi: 10.1016/0005-2736(78)90440-6. [DOI] [PubMed] [Google Scholar]
- Kunkel S. L., Robertson D. C. Purification and chemical characterization of the heat-labile enterotoxin produced by enterotoxigenic Escherichia coli. Infect Immun. 1979 Aug;25(2):586–596. doi: 10.1128/iai.25.2.586-596.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Mekalanos J. J., Collier R. J., Romig W. R. Purification of cholera toxin and its subunits: new methods of preparation and the use of hypertoxinogenic mutants. Infect Immun. 1978 May;20(2):552–558. doi: 10.1128/iai.20.2.552-558.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mug-Opstelten D., Witholt B. Preferential release of new outer membrane fragments by exponentially growing Escherichia coli. Biochim Biophys Acta. 1978 Apr 4;508(2):287–295. doi: 10.1016/0005-2736(78)90331-0. [DOI] [PubMed] [Google Scholar]
- Nagy B., Moon H. W., Isaacson R. E. Colonization of porcine intestine by enterotoxigenic Escherichia coli: selection of piliated forms in vivo, adhesion of piliated forms to epithelial cells in vitro, and incidence of a pilus antigen among porcine enteropathogenic E. coli. Infect Immun. 1977 Apr;16(1):344–352. doi: 10.1128/iai.16.1.344-352.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nagy B., Moon H. W., Isaacson R. E. Colonization of porcine small intestine by Escherichia coli: ileal colonization and adhesion by pig enteropathogens that lack K88 antigen and by some acapsular mutants. Infect Immun. 1976 Apr;13(4):1214–1220. doi: 10.1128/iai.13.4.1214-1220.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmitz J., Preiser H., Maestracci D., Ghosh B. K., Cerda J. J., Crane R. K. Purification of the human intestinal brush border membrane. Biochim Biophys Acta. 1973 Sep 27;323(1):98–112. doi: 10.1016/0005-2736(73)90434-3. [DOI] [PubMed] [Google Scholar]
- Sellwood R., Gibbons R. A., Jones G. W., Rutter J. M. Adhesion of enteropathogenic Escherichia coli to pig intestinal brush borders: the existence of two pig phenotypes. J Med Microbiol. 1975 Aug;8(3):405–411. doi: 10.1099/00222615-8-3-405. [DOI] [PubMed] [Google Scholar]
- Smith W. P., Tai P. C., Thompson R. C., Davis B. D. Extracellular labeling of nascent polypeptides traversing the membrane of Escherichia coli. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2830–2834. doi: 10.1073/pnas.74.7.2830. [DOI] [PMC free article] [PubMed] [Google Scholar]
- So M., Dallas W. S., Falkow S. Characterization of an Escherichia coli plasmid encoding for synthesis of heat-labile toxin: molecular cloning of the toxin determinant. Infect Immun. 1978 Aug;21(2):405–411. doi: 10.1128/iai.21.2.405-411.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stirm S., Orskov F., Orskov I., Mansa B. Episome-carried surface antigen K88 of Escherichia coli. II. Isolation and chemical analysis. J Bacteriol. 1967 Feb;93(2):731–739. doi: 10.1128/jb.93.2.731-739.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
- Wensink J., Gankema H., Jansen W. H., Guinée P. A., Witholt B. Isolation of the membranes of an enterotoxigenic strain of Escherichia coli and distribution of enterotoxin activity in different subcellular fractions. Biochim Biophys Acta. 1978 Dec 4;514(1):128–136. doi: 10.1016/0005-2736(78)90082-2. [DOI] [PubMed] [Google Scholar]
- Witholt B., Boekhout M., Brock M., Kingma J., Heerikhuizen H. V., Leij L. D. An efficient and reproducible procedure for the formation of spheroplasts from variously grown Escherichia coli. Anal Biochem. 1976 Jul;74(1):160–170. doi: 10.1016/0003-2697(76)90320-1. [DOI] [PubMed] [Google Scholar]
- de Leij L., Kingma J., Witholt B. Insertion of newly synthesized proteins into the outer membrane of Escherichia coli. Biochim Biophys Acta. 1978 Sep 22;512(2):365–376. doi: 10.1016/0005-2736(78)90260-2. [DOI] [PubMed] [Google Scholar]
- de Leij L., Kingma J., Witholt B. Nature of the regions involved in the insertion of newly synthesized protein into the outer membrane of Escherichia coli. Biochim Biophys Acta. 1979 May 17;553(2):224–234. doi: 10.1016/0005-2736(79)90227-x. [DOI] [PubMed] [Google Scholar]