Evidence for Covalent Attachment of Diphytanylglyceryl Phosphate to the Cell-surface Glycoprotein of Halobacterium halobium
| Autor: | Akihiro Kikuchi, Hiroshi Sagami, Kyozo Ogura |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Halobacterium salinarum
Molecular Sequence Data Mevalonic Acid Halobacterium Biochemistry Mass Spectrometry Hydrolysate chemistry.chemical_compound Dolichol Polyisoprenyl Phosphates medicine Trypsin Amino Acid Sequence Molecular Biology chemistry.chemical_classification Gel electrophoresis Membrane Glycoproteins Molecular Structure biology Acid phosphatase Cell Biology biology.organism_classification Amino acid chemistry biology.protein Chromatography Thin Layer Glycoprotein Bacterial Outer Membrane Proteins medicine.drug |
| Zdroj: | Journal of Biological Chemistry. 274:18011-18016 |
| ISSN: | 0021-9258 |
| Popis: | In a previous study, we demonstrated the occurrence of novel proteins modified with a diphytanylglyceryl group in thioether linkage in Halobacterium halobium (Sagami, H., Kikuchi, A., and Ogura, K. (1995) J. Biol. Chem. 270, 14851–14854). In this study, we further investigated protein isoprenoid modification in this halobacterium using several radioactive tracers such as [3H]geranylgeranyl diphosphate. One of the radioactive bands observed on SDS-polyacrylamide gel electrophoresis corresponded to a periodic acid-Schiff stain-positive protein (200 kDa). Radioactive and periodic acid-Schiff stain-positive peptides (28 kDa) were obtained by trypsin digestion of the labeled proteins. The radioactive materials released by acid treatment of the peptides showed a similar mobility to dolichyl (C55) phosphate on a normal-phase thin-layer plate. However, radioactive hydrolysates obtained by acid phosphatase treatment co-migrated not with dolichol (C55–65), but with diphytanylglycerol on both reverse- and normal-phase thin-layer plates. The mass spectrum of the hydrolysate was also coincident with that of diphytanylglycerol. The partial amino acid sequences of the 28-kDa peptides were found in a fragment (amino acids 731–816) obtainable by trypsin cleavage of the known cell-surface glycoprotein of this halobacterium. These results indicate that the cell-surface glycoprotein (200 kDa) is modified with diphytanylglyceryl phosphate. |
| Databáze: | OpenAIRE |
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