Abstract
To obtain small membrane markers easily accessible to the charged groups of the cell surface, we prepared, from hemeundecapeptide (HUP), three derivatives that maintain the peroxidatic activity: the anionized hemeundecapeptide, Mr 1,963, estimated diameter 1.68 nm, pl 3.5, for the detection of basic groups; and both a cationized hemeundecapeptide containing predominantly tertiary amino groups, Mr 2,215, estimated diameter 1.75 nm, pl 9.0, and a cationized hemeundecapeptide containing only primary amino groups, Mr 2,271, estimated diameter 1.75 nm, pl 10.6, for labeling acidic residues. The markers were perfused in situ in mice to label the luminal surface of fenestrated endothelium of pancreatic capillaries. Specimens were processed through the cytochemical reaction for peroxidatic activity and examined by electron microscopy. The anionized HUP and HUP (pl 4.85) marked the plasmalemma proper, the coated pits, and the membrane and diaphragms of plasmalemmal vesicles and transendothelial channels. The cationized HUP containing predominantly tertiary amino groups (pl 9.0) decorated all cell surface components with the exception of plasmalemmal vesicles and channels; the latter were, however, labeled by the cationized HUP containing only primary groups (pl 10.6), which suggests that these structures contain on their luminal surface very weak acidic residues of high pKa values. The fact that the membrane of plasmalemmal vesicles can discriminate against permeant cationic macromolecules only up to a pl of approximately 9.0 indicates that in the electrostatic restriction there is a charge limit. In the case of fenestrated capillary endothelium, the upper charge limit seems to be a pl of approximately 9.0. In these vessels, the charge discrimination is effective for molecules as small as 2 nm.
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- Danon D., Goldstein L., Marikovsky Y., Skutelsky E. Use of cationized ferritin as a label of negative charges on cell surfaces. J Ultrastruct Res. 1972 Mar;38(5):500–510. doi: 10.1016/0022-5320(72)90087-1. [DOI] [PubMed] [Google Scholar]
- Donath E., Gingell D. A sharp cell surface conformational transition at low ionic strength changes the nature of the adhesion of enzyme-treated red blood cells to a hydrocarbon interface. J Cell Sci. 1983 Sep;63:113–124. doi: 10.1242/jcs.63.1.113. [DOI] [PubMed] [Google Scholar]
- Feder N. Microperoxidase. An ultrastructural tracer of low molecular weight. J Cell Biol. 1971 Oct;51(1):339–343. doi: 10.1083/jcb.51.1.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ghitescu L., Fixman A. Surface charge distribution on the endothelial cell of liver sinusoids. J Cell Biol. 1984 Aug;99(2):639–647. doi: 10.1083/jcb.99.2.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
- Herzog V., Fahimi H. D. A new sensitive colorimetric assay for peroxidase using 3,3'-diaminobenzidine as hydrogen donor. Anal Biochem. 1973 Oct;55(2):554–562. doi: 10.1016/0003-2697(73)90144-9. [DOI] [PubMed] [Google Scholar]
- Plattner H., Wachter E., Gröbner P. A heme-nonapeptide tracer for electron microscopy. Preparation, characterization and comparison with other heme-tracers. Histochemistry. 1977 Aug 22;53(3):223–242. doi: 10.1007/BF00511078. [DOI] [PubMed] [Google Scholar]
- Schodt K. P., Gelman R. A., Blackwell J. The effect of changes in salt concentration and pH on the interaction between glycosaminoglycans and cationic polypeptides. Biopolymers. 1976 Oct;15(10):1965–1977. doi: 10.1002/bip.1976.360151008. [DOI] [PubMed] [Google Scholar]
- Simionescu M., Simionescu N., Palade G. E. Preferential distribution of anionic sites on the basement membrane and the abluminal aspect of the endothelium in fenestrated capillaries. J Cell Biol. 1982 Nov;95(2 Pt 1):425–434. doi: 10.1083/jcb.95.2.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simionescu M., Simionescu N., Silbert J. E., Palade G. E. Differentiated microdomains on the luminal surface of the capillary endothelium. II. Partial characterization of their anionic sites. J Cell Biol. 1981 Sep;90(3):614–621. doi: 10.1083/jcb.90.3.614. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simionescu N., Siminoescu M., Palade G. E. Permeability of muscle capillaries to small heme-peptides. Evidence for the existence of patent transendothelial channels. J Cell Biol. 1975 Mar;64(3):586–607. doi: 10.1083/jcb.64.3.586. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simionescu N., Simionescu M., Palade G. E. Differentiated microdomains on the luminal surface of the capillary endothelium. I. Preferential distribution of anionic sites. J Cell Biol. 1981 Sep;90(3):605–613. doi: 10.1083/jcb.90.3.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Turner M. R., Clough G., Michel C. C. The effects of cationised ferritin and native ferritin upon the filtration coefficient of single frog capillaries. Evidence that proteins in the endothelial cell coat influence permeability. Microvasc Res. 1983 Mar;25(2):205–222. doi: 10.1016/0026-2862(83)90016-x. [DOI] [PubMed] [Google Scholar]
- Wissig S. L., Williams M. C. Permeability of muscle capillaries to microperoxidase. J Cell Biol. 1978 Feb;76(2):341–359. doi: 10.1083/jcb.76.2.341. [DOI] [PMC free article] [PubMed] [Google Scholar]