Resorufin-α-D-mannopyranoside

Product ID: M1340



Unit SizePriceQuantity 
5mg
$157.69
  • Buy 5 for $126.15 each and save 21%

Availability: In stock


Description

Red fluorogenic substrate can be used to monitor intracellular mannosidase activity using a continuous assay format. Golgi apparatus function and health as well as certain diseases(mannosidosis) can be determined.

Recent interest in processing enzymes in the ER and Golgi has been initiated to screen for treatment options in a variety of genetic enzymatic defects that lead to phenotypic lysosomal storage diseases such as alpha-mannosidosis. In particular, the human deficiency in Golgi alpha-mannosidase II is characterized by congenital dyserythropoietic anemia with splenomegaly and various additional abnormalities and complications. This substrate produces the red fluorophore resorufin that can be monitored using EX 550 nm and EM at 595 nm. Because the pKa of resorufin is 5.8, it allows continuous measurement of fluorescence turnover at or near physiological pH values found in acidic organelles such as lysosomes or the Golgi. The Golgi Mannosidase II enzyme operates at a pH optimum of pH 5.75, while the lysosomal mannosidase enzyme has a pH optimum of 4.5. Our results indicate that this substrate is very useful in measuring Golgi a-Man II activity. Resorufin alpha-D-mannoside also exhibited greater activity towards Golgi Mannosidase II versus human lysosomal mannosidase, indicating it may be suitable for specific intracellular Golgi staining and activity analysis. Use of this new substrate in screening new therapeutics for alpha-mannosidosis or for use in inhibiting transport and secretion of peptides and proteins from cells is possible. Excitation/Emission data is for the enzymatic product, resorufin.

Technical Data
SKU M1340
Unit Size 5mg
Absorption 571nm
Emission Wavelength 595
Detection Method Fluorescence
Molecular Formula C₁₈H₁₇NO₈
Molecular Weight 375.33
Storage Conditions -20C, Desiccated

References and Citations

Citations:

  • Walker AS, Rablen PX, Schepart A. (2016) "Rotamer-Restricted Fluorogenicity of the Bis-Arsenical ReAsH." J. Am. Chem. Soc. DOI: 10.1021/jacs.6b03422.
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    References:

    • Coleman DJ, Kuntz DA, Venkatesan M, Cook GM, Williamson SP, Rose DR, Naleway JJ. (2010) "A long-wavelength fluorescent substrate for continuous fluorometric determination of alpha-mannosidase activity: resorufin alpha-D-mannopyranoside." Anal Biochem. 399(1):7-12.
    • Naleway JJ, Coleman DJ, Cook GM, Kuntz D, Williamson SP, Sim L, Venkatesan M, Rose DR, (2008) "New Resorufin-Based Glycosidase Substrates for Use in Continuous Analysis of Glycosidase Activity in Acidic Organelles." presented at the XXIVth International Carbohydrate Symposium, Oslo, Norway, July, 2008.
    • Coleman DJ, Studler MJ, Naleway JJ (2007) "A long-wavelength fluorescent substrate for continuous fluorometric determination of cellulase activity: resorufin-beta-D-cellobioside." Anal. Biochem. 371: 146-153.
    • Moreman KW, (2002) "Golgi alpha-mannosidase II deficiency in vertebrate systems: implications for asparagine-linked oligosaccharide processing in mammals." Biochim. Biophys. Acta 1573(3): 225-235.
    • Li B, Kawatkar SP, George S, Strachan H, Woods RJ, Siriwardena A, Moremen KW, Boons GJ, (2004) "Inhibition of Golgi Mannosidase II with Mannostatin A Analogues: Synthesis, Biological Evaluation, and Structure-Activity Relationship Studies." ChemBioChem 5(9):1220-1227.
    • Moremen KW, Nairn A. (2013) Chapter: "Mannosidase, alpha, class 2a1 (MAN2A1, Golgi α-mannosidase II)" Handbook of Glycosyltransferases and Related Genes, Edition: 2nd; Publisher: Springer-Verlag, Editors: N. Taniguchi, K. Honke, M. Fukuda, H. Narimatsu, Y. Yamaguchi, T. Angata, pp.1313-1326.
    • Pomorski A, Adamczyk J, Bishop AC, Krezel A. (2015) "Probing the target-specific inhibition of sensitized protein tyrosine phosphatases with biarsenical probes." Org & Biomol Chem 13:1395-1403.
    • Cheng X, Hiras J, Deng K, Northen T. (2013) "High throughput nanostructure-initiator mass spectrometry screening of microbial growth conditions for maximal β-glucosidase production." Frontiers in Microbiology 4:365.
    • Reindl W, Deng K, Gladden JM, Northen T. (2011) "Colloid-based multiplexed screening for plant biomass-degrading glycoside hydrolase activities in microbial communities." Energy & Environ Sci 4(8):2884-2893.
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