Purification and characterization of an isoform of crustacean hyperglycemic hormone from the eyestalk ofMacrobrachium rosenbergii

✍ Scribed by Sithigorngul, Weerawan; Jaideechoey, Sompong; Saraithongkum, Wanpen; Longyant, Siwaporn; Sithigorngul, Paisarn

• Book ID: 101228639
• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 151 KB
• Volume: 284
• Category: Article
• ISSN: 0022-104X
• DOI: 10.1002/(sici)1097-010x(19990701)284:2<217::aid-jez11>3.0.co;2-5

✦ Synopsis

Isolation of the crustacean hyperglycemic hormone (CHH) from the eyestalk of the giant freshwater prawn Macrobrachium rosenbergii was performed using 5,000 ground eyestalks extracted in methanol-acetic acid-water (90:1:9). After the extract was partially purified using C18 cartridges, it was further purified by eight steps of RP-HPLC using four kinds of columns: C18, C8, cyano and phenyl, and three solvent systems: acetonitrile (ACN)/trifluoroacetic acid, ACN/heptafluoroacetic acid and ACN/triethylammonium acetate. The bioassay of CHH during purification was done by injection of eluate fractions into eyestalk-ablated prawns and determination of the ability of the fractions to elevate glucose in the haemolymph. A complete amino acid sequence analysis was performed on one isoform of CHH (Mar-CHH-1), consisting of 71 residues. The sequence of Mar-CHH-1 shows considerable similarity (45-68%) to CHHs reported in other crustaceans. It is expected that there might be more than one isoform of CHH in M.