Efficient photoaffinity labeling of human beta-hexosaminidase A. Synthesis and application of 3-azi-1-[(2-acetamido-2-deoxy-1-beta- D-glucopyranosyl)thio]- and -galactopyranosyl)thio]butane.
Two photolabile thioglycosides (8 and 9) were synthesized by Koenigs-Knorr type glycosylation. These compounds, being enzyme-resistant analogues of N-acetylhexosaminides, were shown to be good competitive inhibitors of lysosomal beta-hexosaminidase (2-acetamido-2-deoxy- beta-D-hexoside acetamidodeoxyhexohydrolase, EC 3.2.1.52) action. For photoaffinity labeling 3H-labeled 8a was prepared by enzymatic oxidation with galactose oxidase followed by reduction with sodium [3H]borohydride. Compound 8a, when photolyzed in the presence of hexosaminidase, specifically labeled both subunits of the enzyme.
Kuhn
C S
CS
Institut für Organische Chemie und Biochemie, Universität Freiburg, Germany.
Lehmann
J
J
Sandhoff
K
K
D000345:Affinity Labels / Q000138:chemical synthesis*; D001391:Azo Compounds / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D001894:Borohydrides; D006801:Humans; D007202:Indicators and Reagents; D008247:Lysosomes / Q000201:enzymology; D015394:Molecular Structure; D011485:Protein Binding; D013329:Structure-Activity Relationship; D013865:Thioglycosides / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D014316:Tritium; D001619:beta-N-Acetylhexosaminidases / Q000037:antagonists & inhibitors*
D000345:Affinity Labels; D001391:Azo Compounds; D001894:Borohydrides; D007202:Indicators and Reagents; D013865:Thioglycosides; D014316:Tritium; C076591:3-azi-1-((2-acetamido-2-deoxy-1-galactopyranosyl)thio)butane; C076589:3-azi-1-((2-acetamido-2-deoxy-1-glucopyranosyl)thio)butane; D001619:beta-N-Acetylhexosaminidases