N‐Glycan structures of squid rhodopsin

Abstract

To determine the glycoforms of squid rhodopsin, N‐glycans were released by glycoamidase A digestion, reductively aminated with 2‐aminopyridine, and then subjected to 2D HPLC analysis [Takahashi, N., Nakagawa, H., Fujikawa, K., Kawamura, Y. & Tomiya, N. (1995) Anal. Biochem. 226 , 139–146]. The major glycans of squid rhodopsin were shown to possess the α1–3 and α1–6 difucosylated innermost GlcNAc residue found in glycoproteins produced by insects and helminths. By combined use of 2D HPLC, electrospray ionization‐mass spectrometry and permethylation and gas chromatography‐electron ionization mass spectrometry analyses, it was revealed that most (85%) of the N‐glycans exhibit the novel structure Manα1–6(Manα1–3)Manβ1–4GlcNAcβ1–4(Galβ1–4Fucα1–6)(Fucα1–3)GlcNAc.