Design and synthesis of substrate and internal standard conjugates for profiling enzyme activity in the Sanfilippo syndrome by affinity chromatography/electrospray ionization mass spectrometry

We describe the design and synthesis of substrate and internal standard con jugates for application in profiling enzyme activity of the enzymes alpha - D-2-deoxy-2-N-sulfonamido-glucosamine sulfamidase, alpha -D-2-deoxy-2-N-ace tyl-glucosamine hydrolase, acetyl-coenzymeA: alpha -D-2-deoxy-2 -amino-gluc osamine transferase, and alpha -D-2-deoxy-2-N-acetyl-glucosamine-6-sulfate sulfatase. Deficiency of any one of these enzymes results in a single clini cal phenotype known as Sanfilippo syndrome. Such substrates have been prove n effective in the confirmation of enzyme deficiency by a combination of af finity chromatography (AC) and electrospray ionization mass spectrometry (E SIMS), which forms the foundation for a new analytical technology (ACESIMS) of general interest and application to clinical and biomedical research.