P-Nitrophenyl a-D-mannopyranoside

T. Nishio, Y. Miyake, H. Tsujii, W. Hakamata, K. Kadokura, and T. Oku, Hydrolytic activity of a-mannosidase against deoxy derivatives of p-nitrophenyl a-D-mannopyranoside, Biosci. Biotechnol. Biochem., 60 (1996) 2038-2042. 

Enzymatic mannosylation of the alkaloids by a-mannosidase from Canavalia ensiformis (Jack beans) was accomplished via two different strategies - transglycosylation using p-nitrophenyl-a-D-mannopyranoside or reversed glycosylation using a high concentration of mannose [46] (Fig. 10). 

For the assay of a-D-mannosidase, the incubation mixture employed in our laboratory contained 0.5 ml of M acetate buffer at a pH appropriate for the particular enzyme preparation, 1.5 ml of 16 mM p-nitrophenyl a-D-mannopyranoside, 1.5 ml of water (which could be replaced by other additives as required), and 0.5 ml of suitably diluted, enzyme preparation. After 1 hour at 37°, the reaction was terminated, and the color was developed by adding 4 ml of 0.4 M glycine-sodium hydroxide buffer, pH 10.5. The mixture was centrifuged for 15 minutes at 1500 g, and the color intensity of the liberated p-nitrophenol (25-150 fig) in the supernatant liquor was measured on a Spekker photoelectric absorptiometer, with use of Ilford No. 601 violet filters having maximal transmission at 430 nm, and a 1-cm light path. Separate control-experiments for enzyme and sub-... 

D-Mannono-1,5-lactone68 gave a value of 0.07 mM for Kj with rat-epididymal a-D-mannosidase, compared with 12 mM for Km for p-nitrophenyl a-D-mannopyranoside,88 and 57 mM for phenyl a-D-mannopyranoside.65 With jack-bean a-D-mannosidase, the values... 

As discussed under Purification and pH and Stability [see Sections 11,4 (p. 409) and 11,6 (p. 413)], a-D-mannosidase is unstable at low pH values unless Zn2+ is added. In the following experiments employing a partially purified, jack-bean meal preparation (see Table V, stage 3 p. 410) to which Zn2+ had not been added, the enzyme was pre-incubated at 37° and pH 5 before assay at the same pH with p-nitrophenyl a-D-mannopyranoside as the sub-... 

The fraction of protein, eluted from Sephadex G-75 which behaves as a single homogeneous protein with respect to its interactions with M2 was also studied with regard to its interaction with monosaccharides. The association constant of the interaction of M2 with intact con A is 19 times greater than that obtained for the interaction of p-nitrophenyl a-D-mannopyranoside with con A. The normalized spectra (Figure 7) of the monosaccharide and disaccharide complexes were found to be very similar suggesting that the interactions between con A and the nitrophenyl group which causes the spectral perturbation are the same for both complexes. 

Activation Parameters for Formation and Breakdown of Complexes of Con A with M2 and With p-Nitrophenyl a-D-Mannopyranoside... 

R. C. Gasman and D. C. Johnson, C-2 Oxyanion participation in the base-catalyzed cleavage of p-nitrophenyl 3-D-galactopyranoside and p-nitrophenyl a-D-mannopyranoside, J. Org. Chem., 31 (1966) 1808-1830. 

Firon et al. (30) found that the maimose binding sites of E. coli. is a hydrophobic region. Therefore, mannose with hydrophobic linker, such as p-nitrophenyl-a-D-mannopyranoside, will exibit stronger binding. We designed two different linker for mannose a PEG linker (hydrophilic, such as 16A) and p-amidophen l linker (hydrophobic, such as 16B). 

Wade et al. [46] have compared the experimental band profiles of p-nitrophenyl-ff-D-mannopyranoside on silica-bonded Concanavalin A, obtained in affinity chromatography, and the best fit parameters to their model. This model i.e., Thomas model) uses a Langmuir kinetic and neglects the axial dispersion. The best values of the parameters are calculated using a Simplex program to minimize the sum of the residuals of the predided and experimental band profiles. Figure 14.10 illustrates the results obtained and shows excellent agreement. 

Diethyl azodicarboxylate and triphenylphosphine permit the condensation between aldose derivatives with a free reducing centre and phenols. 2,3 5,6-Di-0-isopropylidene-D-mannose gave 67% of the phenyl o-D-mannofuranoside derivative and 16% of the /3-anomer, while 2,3 4,6-di-O-cyclohexylidene-a-D-mannose afforded access to p-nitrophenyl /3-D-mannopyranoside in 58% yield and other, substituted aryl analogues. Unsubstituted sugars can be subjected to the procedure, D-glucose giving mainly the phenyl 3-pyranoside. ... 

On treatment with NIS, TfOAg a mixture of the 2,6-tethered 6 and phenyl tetra-O-acetyl-l-thio-a-D-mannopyranoside underwent (l- 6)-cyclization of 6 and slower 3-a-mannosylation of the product, the former step being favoured by its intramolecular character in this one-pot procedure. a-D-Man-(l- 3)-[a-D-Man-(l- 6)]-a-D-ManOMe was consequently made in 30% yield. Alternatively this trisaccharide was made in 42% yield by direct selective mannosylation with the O-acetylated trichloroacetimidate (2 equiv) of p-nitrophenyl a-D-man-nopyranoside. The product was converted to the p-acrylamide glycoside and copolymerized with 2-acrylamido-N,iV-dimethylamine to give a product tested for lectin binding. ... 

Fig. 13.—A Agar diffusion of immune sera (Se), 1-thio-D-mannose antibodies (A ), and anti-BSA antibodies (A2) against Man-S-BSA (/) and BSA (2). B Agar-diffusion plate of anti-Man-S-antibodies and antibodies oxidized by peroxypropanoic acid for 0, 4, and 8 h. C, D, E, and F Hapten inhibition by agar diffusion, A = purified anti-Man-S antibodies I = antibodies + p-nitrophenyl 1 -thio-a-D-mannopyranoside 12 = antibodies + D-mannose I3 = antibodies + ethyl 1-thio-a-o-mannoside 1 to 6, outer wells contain decreasing concentration of Man-S-BSA. (Reprinted with permission from Journal of Protein Chemistry, Volume 9, J. H. Pazur, B. Liu, Nan Q Li, and Y. C. Lee, pp. 143-150, copyright 1990 Journal of Protein Chemistry.)...

Nitrophenyl-a-L-fuco/galacto/rhamno/manno-pyranosides can be made by the reaction of the acetate protected sugar with p-nitrophenol and zinc chloride at 120 °C under vacuum [7]. p-Nitrophenyl-p-D-gluco/galactopyranoside can be generated from the reaction of the acetylated hexopyranosyl bromides with sodium p-nitrophenoxide in DMF [8], followed by deprotection, and /7-nitrophenyl-(3-D-mannopyranoside can be synthesised from mannose via 2,3 4,6-di-0-cyclohex-ylidene-a-D-mannopyranose [9]. 

AA AAm Con A DSS DSS-gel LCST MAPTAC MBA MP MP-gel NIPA ONPG PVMA SSPG Tc TMED acrylic acid acrylamide concanavalin A dextran sulfate sodium gel containing Con A/DSS complex lower critical solution temperature [(methacrylamide)propyl]trimethylammonium chloride /V,/V -methylenebis(acrylamide) a-methyl-D-mannopyranoside gel containing Con A/MP complex /V-isopropylacrylamide O-nitrophenyl-P-D-galactopyranoside poly(vinyl methyl ether) stimulus-sensitive polymer gel transition temperature /V,/V,/V, /V -tetramethylethylenediamine... 

The inhibition of precipitin formation by 1-thio-D-mannose and derivatives is shown in the results recorded in Plates C, D, E, and F of Fig. 13. The sulfur derivatives, p-nitrophenyl 1-thio-D-mannopyranoside (well Ii) and ethyl 1-thio-D-mannopyranoside (well I3), caused a marked decrease in the amount of precipitin formation, in comparison to that obtained with the native antibodies (Well A,). However, mannose did not decrease the amount of precipitin formation (well I2). This compound did not bind to the combining site of the antibody. Apparently, the thio group at position 1 of the mannose is required for the binding to occur. 

Nitrophenyl mannoside a-D-P)ranose-/orm, N-73 4-Nitrophenyl mannoside p-D-Pyranose-/orm, N-73 l,2,3A6-Penta-0-acetyl-a-D-mannopyranose, M-114 l,2,3A6-Penta-0-acetyl-p-D-mannopyranose, M-114 l,2,3A6-Penta-0-benzoyl-p-D-mannopyranose, M-114 4-Pentenyl mannopyranoside a-D-form, P-34 4-Pentenyl mannopyranoside p-D-/orm, P-34... 

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