Mass Spectrometric Characterisation of Post-Translational Modification and Genetic Variation in Human Tetranectin
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Michel Jaquinod
Abstract
Tetranectin, a plasminogen-binding trimeric C-type lectin-like protein primarily involved in tissue remodeling and development, was scanned for covalent modifications and sequence heterogeneity, using a combination of mass spectrometric and classical protein chemical analytical methods. Electrospray ionisation mass spectrometry showed the presence of eight components of different mass and abundance in plasma tetranectin, all of higher mass than that calculated from the cDNA sequence. To identify and locate residues accounting for the heterogeneity, samples of tetranectin were subjected to proteolytic cleavage. Peptide fragments, in mixtures or in purified form, were analysed by matrix-assisted-laser-desorptionionisation mass spectrometry and, where required, by Edman sequencing and compared to the cDNA sequence. Our results show that the mass heterogeneity in plasma tetranectin is due to sequence heterogeneity at position 85 and the presence of a partially sialylated oligosaccharide prosthetic group attached to Thr-4. Residue 85 is encoded in the cDNA as a Ser residue, but plasma tetranectin is a 1:1 mixture of Ser- 85 and Gly-85 sequence variants. Mass spectrometric analysis of enzymatic and mild acid hydrolysates of an N-terminal glycopeptide showed that the composition and partial covalent structure of the O-linked oligosaccharide prosthetic group is ≤ N-acetylhexosamine ≤ [hexose, (sialic acid)0–3].
Copyright © 1999 by Walter de Gruyter GmbH & Co. KG
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Artikel in diesem Heft
- Secretory Immunoglobulin A: from Mucosal Protection to Vaccine Development
- Cloning of karyopherin-α3 from Drosophila Through Its Interaction with the Nuclear Localization Sequence of Germ Cell-Less Protein
- Regulation of Bile Salt Export Pump mRNA Levels by Dexamethasone and Osmolarity in Cultured Rat Hepatocytes
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- Synthesis, Cytotoxicity and Antitumor Activity of Platinum(II) Complexes of Cyclopentanecarboxylic Acid Hydrazide
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- Mass Spectrometric Characterisation of Post-Translational Modification and Genetic Variation in Human Tetranectin
- Complete Amino Acid Sequence Determination of the Major Allergen of Peach (Prunus persica) Pru p1
- Identification of Immunoreactive Tissue Prokallikrein on the Surface Membrane of Human Neutrophils
- An Evaluation of Techniques for the Extraction and Amplification of DNA from Naturally Shed Hairs
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