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标题：Chemical and Immunological Characterization of the Two α-N-Acetylgalactosaminidases from Squid Liver
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作者：Golam Sadik ; Md Harun or Rashid ; Tomoko Itoh-Nashida 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：2009
卷号：32
期号：8
页码：1469-1472
DOI：10.1248/bpb.32.1469
出版社：The Pharmaceutical Society of Japan
摘要：Based on the inherent α-galactosidase activity, squid liver contains two different α- N -acetylgalactosaminidases (α-GalNAcases): α- N -acetylgalactosaminidase I (α-GalNAcase I), which typically exhibits the α-galactosidase activity and α- N -acetylgalactosaminidase II (α-GalNAcase II), which is devoid of such activity. The molecular properties of the α-GalNAcases that may account for their enzymological differences are as yet unknown. In this study, we have characterized and compared the chemical and immunological properties of α-GalNAcase I and α-GalNAcase II. Analysis of the N-terminal sequence of the first twenty amino acids revealed the striking homology between α-GalNAcase I and α-GalNAcase II. Digestion of α-GalNAcase I and α-GalNAcase II generated the peptide maps that display similarities in peptide pattern, indicating their close relationship in structure. Polyclonal antibodies were generated in rabbits against the purified α-GalNAcase I and α-GalNAcase II for comparison of the immunological properties. Both Western blot and surface plasmon resonance (SPR) studies showed that the anti-α-GalNAcase II antibody reacted with both α-GalNAcase I and α-GalNAcase II, whereas the anti-α-GalNAcase I antibody reacted only with α-GalNAcase I, indicating the presence of common as well as unique antigenic determinants on α-GalNAcase I and α-GalNAcase II. Taken together, these results suggest that α-GalNAcase I and α-GalNAcase II are closely related with regard to structure and that their nonhomologous domains are possibly responsible for the differences in enzymatic properties.
关键词：squid liver;α-N-acetylgalactosaminidase;immunological property;N-terminal amino acid sequence;peptide map