Journal of comparative physiology

, Volume 153, Issue 2, pp 207–221 | Cite as

Peptides in the sinus gland ofCardisoma carnifex: isolation and amino acid analysis

  • Robert W. Newcomb
Article

Summary

Reversed phase liquid chromatography (RPLC) has been used to isolate most of the peptides in the sinus gland of the land crabCardisoma carnifex. The peptides accounted for at least 50% (5–10 μg) of the non-amino acid primary amine compounds of the sinus gland. Peptides were characterized by (1) amino acid analysis; (2) bioassay for erythrophore concentrating, melanophore dispersing and hyperglycemic activity; (3) a comparison of absorption at 280 nm with intrinsic fluorescence; (4) RPLC elution properties relative to the shrimp crythrophore concentrating hormone (ECH); and (5) post-column detection with the fluorescamine reagent. High sensitivity peptide maps were obtained by RPLC of the peptide fluoreseamine derivatives.

Peptides in theCardisoma sinus gland can be summarized as:
  1. 1.

    Relatively hydrophobic peptides which elute in three broad peaks, labeled J, K, and L, each of which is possibly heterogeneous. The amino acid compositions of these fractions are similar to those previously reported by others for the hyperglycemic hormones of several species and at least two possessed this activity.

     
  2. 2.

    Compounds which eluted before the J, K and L peptide fractions, and which both absorbed light at 280 nm and possessed intrinsic fluorescence, Because of these properties, these compounds were considered likely to be peptides. One is indistinguishable from the shrimp ECH by (a) chromatography on two separate columns, (b) amino acid analysis, (c) the ratio of absorbance at 280 nm to intrinsic fluorescence, and (d) bioactivity on the erythrophores ofOcypode pallidula.

     
  3. 3.

    Peptides (all novel, labeled C-I) which did not possess tyr, trp, val, ile or lys, and which were more polar than the J, K and L peptide fractions. By amino acid analysis, all of these could be explained by cleavage from the largest, H, of M.W. 3300 to 4000. On a mole basis, the C-I peptides accounted for almost all of the small (M.W. 500–4000) peptide material of theCardisoma sinus gland.

     

Keywords

Peptide Primary Amine Amino Acid Analysis Intrinsic Fluorescence Reverse Phase Liquid Chromatography 

Abbreviations

CHH

crustacean hyperglycemic hormone

ECH

erythrophore concentrating hormone

ODS

octadecyl silica

OPA

o-phthaldialdehyde

PDH

pigment dispersing hormone

RPLC

reversed phase high pressure liquid chromatography

TFA

trifluoroacetic acid

THF

tetrahydrofuran

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Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • Robert W. Newcomb
    • 1
  1. 1.Department of Biochemistry and Biophysics and Békésy Laboratory of NeurobiologyUniversity of HawaiiHonoluluUSA

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