Determination of the covalent structure of an N- and C-terminally blocked glycoprotein from endocuticle of Locusta migratoria. Combined use of plasma desorption mass spectrometry and Edman degradation to study post-translationally modified proteins.
The complete structure of protein isolated from endocuticle of sexually mature locusts, Locusta migratoria, has been determined by a combination of automatic Edman degradation and plasma desorption mass spectrometry. The protein is extensively post-translationally modified. The N-terminal is 5-oxoproline (pyroglutamic acid) and the C-terminal proline residue is amidated. Furthermore, the protein is glycosylated by a single N-acetyl-galactosamine residue at one, two or three threonines. The N-terminal sequence was obtained by analysing the N-acetylated N,O-permethylated derivative using plasma desorption mass spectrometry. The position and type of carbohydrate were determined by combining an HPLC-based carbohydrate analysis with the peak pattern of the phenylthiohydantoin derivative in automatic sequencing and with mass information on peptides. The protein has pronounced similarity to cuticular proteins from larvae of diptera and lepidoptera, but only slight resemblance to the previously sequenced locust exocuticular proteins. This indicates a similarity between soft larval cuticles and locust endocuticle, a similarity which may extend to their mechanical properties.
Talbo
G
G
Department of Molecular Biology, University of Odense, Denmark.
Højrup
P
P
Rahbek-Nielsen
H
H
Andersen
S O
SO
Roepstorff
P
P
D000116:Acetylgalactosamine / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001322:Autoanalysis; D004175:Diptera / Q000235:genetics; D029721:Drosophila Proteins; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics; D006031:Glycosylation; D006110:Grasshoppers / Q000235:genetics; D018398:Homeodomain Proteins; D006868:Hydrolysis; D007301:Insect Hormones / Q000302:isolation & purification; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D012697:Serine Endopeptidases
C062393:Abd-B proteins, Drosophila; D029721:Drosophila Proteins; D006023:Glycoproteins; D018398:Homeodomain Proteins; D007301:Insect Hormones; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D000116:Acetylgalactosamine