Preparation, identfication, and activity assay of lamprey (lampetra japonica) natural intelectins

J Immunoassay Immunochem. 2015;36(4):368-78. doi: 10.1080/15321819.2014.955583.

Abstract

Intelectins play an important role in innate immune response. In a previous study, lamprey inteletins purified by galactose-Sepharose were inactive and insoluble. Herein, we provided a simple and effective method to purify natural intelectins from the serum of lamprey (Lethenteron japonicum) using proteinG agarose. SDS-PAGE, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and mass spectrometry (MS) were used to analyze the purified proteins. The purified proteins were identified to be lamprey serum lectin and intelectinB. The activity analysis results indicated that the proteins had certain extent agglutination activity. The effective method will be useful to study their immune functions and molecular mechanisms.

Keywords: immunoprecipitation; innate immune; intelectin; lamprey; mass spectrometry; purification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agglutination Tests
  • Animals
  • Cytokines* / blood
  • Cytokines* / pharmacology
  • Electrophoresis, Gel, Two-Dimensional
  • Erythrocytes / drug effects
  • Escherichia coli / drug effects
  • Fish Proteins* / blood
  • Fish Proteins* / pharmacology
  • GPI-Linked Proteins* / blood
  • GPI-Linked Proteins* / pharmacology
  • Lampreys / metabolism
  • Lectins* / blood
  • Lectins* / pharmacology
  • Rabbits
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Cytokines
  • Fish Proteins
  • GPI-Linked Proteins
  • Lectins