A chromophore-containing agglutinin from Haliclona manglaris: purification and biochemical characterization

Int J Biol Macromol. 2015 Jan:72:1368-75. doi: 10.1016/j.ijbiomac.2014.10.003.

Abstract

A new chromophore-containing agglutinin (Haliclona manglaris agglutinin (HMA)) was isolated from the tropical sponge H. manglaris. HMA was purified by a combination of hydrophobic interaction chromatography and ion exchange chromatography. Native HMA is a heterotrimer formed by two β-chains (15 kDa) and one α-chain (22 kDa). HMA is a glycoprotein and possesses three intrachain disulfide bonds. Hemagglutinating activity of HMA was stable at neutral pH and temperatures up to 60 °C. HMA was only inhibited by thyroglobulin. Mass spectrometry sequencing and Edman degradation revealed a unique amino acid sequence of about 30%. Moreover, HMA has an organic chromophore of 581 Da, and this characteristic seems to be important to its antioxidant activity. Interestingly, while HMA showed no toxicity against Artemia nauplii and was unable to agglutinate bacterial cells, it did show a high capacity to protect β-carotene against oxidation. Thus, our findings suggest the putative involvement of HMA in the protection of the sponge against oxidation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agglutinins / chemistry*
  • Agglutinins / isolation & purification*
  • Amino Acid Sequence
  • Animals
  • Antioxidants / pharmacology
  • Artemia / drug effects
  • Carbohydrates / analysis
  • Cations, Divalent / pharmacology
  • Chromatography, Gel
  • Chromatography, Reverse-Phase
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Dyes / chemistry*
  • Haliclona / chemistry*
  • Hemagglutination / drug effects
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Molecular Weight
  • Peptides / chemistry
  • Rabbits
  • Sequence Analysis, Protein
  • Sulfhydryl Compounds / chemistry
  • Tandem Mass Spectrometry
  • Temperature

Substances

  • Agglutinins
  • Antioxidants
  • Carbohydrates
  • Cations, Divalent
  • Fluorescent Dyes
  • Peptides
  • Sulfhydryl Compounds