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101.
102.
Studies on the binding site of the galactose-specific agglutinin PA-IL from Pseudomonas aeruginosa 总被引:1,自引:0,他引:1
The binding properties of Pseudomonas aeruginosa agglutinin-I (PA-IL) with
glycoproteins (gps) and polysaccharides were studied by both the
biotin/avidin-mediated microtiter plate lectin-binding assay and the
inhibition of agglutinin-glycan interaction with sugar ligands. Among 36
glycans tested for binding, PA-IL reacted best with two glycoproteins
containing Galalpha1-->4Gal determinants and a human blood group ABO
precursor equivalent gp, but this lectin reacted weakly or not at all with
A and H active gps or sialylated gps. Among the mammalian disaccharides
tested by the inhibition assay, the human blood group Pkactive
Galalpha1-->4Gal, was the best. It was 7.4-fold less active than
melibiose (Galalpha1-->6Glc). PA-IL has a preference for the
alpha-anomer in decreasing order as follows: Galalpha1-->6
>Galalpha1-->4 >Galalpha1-->3. Of the monosaccharides studied,
the phenylbeta derivatives of Gal were much better inhibitors than the
methylbeta derivative, while only an insignificant difference was found
between the Galalpha anomer of methyl- and p -NO2-phenyl derivatives. From
these results, it can be concluded that the combining size of the
agglutinin is as large as a disaccharide of the alpha-anomer of Gal at
nonreducing end and most complementary to Galalpha1-->6Glc. As for the
combining site of PA-IL toward the beta-anomer, the size is assumed to be
less than that of Gal; carbon-6 in the pyranose form is essential, and
hydrophobic interaction is important for binding.
相似文献
103.
Release of spectrin-free vesicles from human erythrocytes during ATP depletion: 1. characterization of spectrin-free vesicles 总被引:1,自引:0,他引:1
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Human erythrocytes incubated without glucose at 37 degrees C (in vitro aging) release spectrin-free vesicles after 12 or more hours. The release of vesicles is dependent upon ATP depletion. If the endogenous level of ATP is maintained, vesicle release is completely inhibited up to 54 h. Vesicle release is independent of hemolysis because in vitro aged cells and cells that maintain their ATP levels lose identical amounts of hemoglobin up to 45 h. 93 percent of all membrane particles released constitute a uniform population of spheres with a diameter of 185 +/- 23nm. These vesicles are of slightly varying densities due to varying contents of hemoglobin. Vesicles contain half the amount of membrane protein that is found in intact membranes when referred to the content of phospholipids phosphorus. This is primarily due to the absence of spectrin. However, their content of protein component III, glycophorin, and cholesterol remains the same as in intact membranes. Thus, the major integral membrane proteins are present in vesicles in similar quantities were surface area as in cells except for the enzyme acetylcholinesterase that is enriched up to twofold. The phospholipids composition of these vesicles is representative of the intact membrane except that the amount of phosphatidic acid is 10-fold higher and the amount of phosphatidylethanolamine is slightly lower than in erythrocytes. These results suggest a selective release of membrane domains that lack peripheral membrane proteins and are enriched in acetylcholinesterase. This release of spectrin-free vesicles from cells aged in vitro could represent an acceleration of the physiological aging process. 相似文献
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