Molecular Weight: 375.33
Storage: F,D
[C=Cold D=Desiccated F=Frozen L=Light Sensitive RT=Room Temperature]
Emission: (in nm) 595
Extinction (103): 550
Molecular Formula: C18H17NO8
CAS Number: [N/A]
Alternative Name: Available Soon
Unit Price (USD):
5 mg $122.00
Bulk Price/unit (when you buy 5 or more): $97.60
Description:
Red fluorogenic substrate can be used to monitor intracellular mannosidase activity using a continuous assay format. Activity has been evaluated for cloned human Golgi α-Mannosidase II as well as human lysosomal α-mannosidase.
Application:
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 ?-mannosidosis or for use in inhibiting transport and secretion of peptides and proteins from cells is possible.
References:
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-?-D-cellobioside." Anal. Biochem. 371: 146-153.
Moreman KW, (2002) "Golgi a-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.
|
