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Characterization of exoskeletal proteins from the American lobster, Homarus americanus
1Department of Molecular Biology, Odense University, Denmark.
Proteins from the calcified exoskeleton of the lobster, Homarus americanus, were extracted and separated by two-dimensional gel-electrophoresis. Electroblotting the proteins onto polyvinylidene difluoride (PVDF) membranes followed by sequence determination gave 16 N-terminal amino-acid sequences and revealed that further eight proteins were N-terminally blocked. The relative molecular mass, M(r), was obtained for most of the electrophoretically separated proteins by means of matrix-assisted laser desorption mass spectrometry (MALDIMS) after electroelution from Coomassie-stained two-dimensional polyacrylamide gels. Eleven proteins were purified from extracts of the exoskeleton by low pressure ion exchange chromatography and reversed-phase high performance chromatography, and their sequences were determined by combined use of Edman degradation and mass spectrometry. Good agreement was obtained between the M(r)-values measured by mass spectrometry and those calculated from the sequences. Five of the sequenced proteins contain two copies of a previously observed 18-residue sequence motif, while a couple of the remaining sequences show similarity to sequences of exoskeletal proteins from shrimps and spiders. Only limited similarity to insect cuticular proteins was observed.
Proteins from the calcified exoskeleton of the lobster, Homarus americanus, were extracted and separated by two-dimensional gel-electrophoresis. Electroblotting the proteins onto polyvinylidene difluoride (PVDF) membranes followed by sequence determination gave 16 N-terminal amino-acid sequences and revealed that further eight proteins were N-terminally blocked. The relative molecular mass, M(r), was obtained for most of the electrophoretically separated proteins by means of matrix-assisted laser desorption mass spectrometry (MALDIMS) after electroelution from Coomassie-stained two-dimensional polyacrylamide gels. Eleven proteins were purified from extracts of the exoskeleton by low pressure ion exchange chromatography and reversed-phase high performance chromatography, and their sequences were determined by combined use of Edman degradation and mass spectrometry. Good agreement was obtained between the M(r)-values measured by mass spectrometry and those calculated from the sequences. Five of the sequenced proteins contain two copies of a previously observed 18-residue sequence motif, while a couple of the remaining sequences show similarity to sequences of exoskeletal proteins from shrimps and spiders. Only limited similarity to insect cuticular proteins was observed.