Nanomolar affinity, iminosugar-based chemical probes for specific labeling of lysosomal glucocerebrosidase.
Three different photoprobes were synthesized to label beta-glucosidases; one probe was based on glucose, two probes on the iminosugar deoxynojirimycin. The affinity of the probes for three different beta-glucosidases was determined. Furthermore, their labeling efficiencies, binding specificities through competition with deoxynojirimycin, and binding specificities in the presence of cell lysate, were evaluated. Especially one showed very high affinity towards non-lysosomal glucoceramidase (IC(50)=20 nM).
van Scherpenzeel
Monique
M
Department of Medicinal Chemistry and Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, PO Box 80082, 3508 TB Utrecht, The Netherlands.
van den Berg
Richard J B H N
RJ
Donker-Koopman
Wilma E
WE
Liskamp
Rob M J
RM
Aerts
Johannes M F G
JM
Overkleeft
Herman S
HS
Pieters
Roland J
RJ
D001419:Bacteria / Q000737:chemistry; D005947:Glucose / Q000737:chemistry / Q000378:metabolism; D005962:Glucosylceramidase / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D050111:Imino Sugars / Q000737:chemistry* / Q000378:metabolism; D008247:Lysosomes / Q000201:enzymology*; D010777:Photochemistry; D011485:Protein Binding
D050111:Imino Sugars; D005962:Glucosylceramidase; D005947:Glucose