Synthesis and Characterization of a Carbene-Generating Biotinylated Lactosylceramide Analog as a Novel Chromogenic Photoprobe for GM3 Synthase

Yasumaru HATANAKA; Makoto HASHIMOTO; Kazuya I.-P. Jwa HIDARI; Yutaka SANAI; Yasuhiro TEZUKA; Yoshitaka NAGAI; Yuichi KANAOKA

doi:10.1248/cpb.44.1111

Synthesis and Characterization of a Carbene-Generating Biotinylated Lactosylceramide Analog as a Novel Chromogenic Photoprobe for GM₃ Synthase

Yasumaru HATANAKA, Makoto HASHIMOTO, Kazuya I.-P. Jwa HIDARI, Yutaka SANAI, Yasuhiro TEZUKA, Yoshitaka NAGAI, Yuichi KANAOKA

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Keywords: photoaffinity labeling, ganglioside, sialyltransferase, biotinylated probe, chromogenic diazirine, avidin-biotin system

JOURNAL FREE ACCESS

1996 Volume 44 Issue 5 Pages 1111-1114

DOI https://doi.org/10.1248/cpb.44.1111

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Abstract

A new biotinylated lactose derivative bearing a nitro-substituted chromogenic diazirine was synthesized. The biotinyl group within the structure enabled the performance of a convenient assay of GM₃ synthase based on avidin-biotin technology, and the K_m values of this biotinylated photoprobe were determined as 40 and 47 μM using bovine brain and rat liver Golgi as enzyme sourced, respectively. Furthermore, the sialylation of lactosylceramide, a natural acceptor substrate for GM₃ synthase, was completitively inhibited by this synthetic analog. The reagent could be a useful chromogenic photoprobe for GM₃ synthase.

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