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Primary structure of human placental 5′-nucleotidase and identification of the glycolipid anchor in the mature form.
- Source :
-
European Journal of Biochemistry . 8/17/90, Vol. 191 Issue 3, p563-569. 7p. - Publication Year :
- 1990
-
Abstract
- A cDNA was cloned coding for human placental 5’-nucleotidase. The 3547-cDNA contains an open reading frame that encodes a 574-residue polypeptide with a calculated size of 63375 Da. The NH2-terminal 26 residues comprises a signal peptide, which is followed by the NH2-terminal sequence of the purified protein. Four potential N-linked glycosylation sites are found in the molecule, accounting for a larger mass of the mature form (71 kDa). The predicted structure contains a hydrophobic amino acid sequence at the COOH terminus, a possible signal for the post-translational modification by glycophosholipid. To confirm this possibility, we tried to isolate and characterize the membrane-anchoring domain of 5’-nucleotidase. BrCN-cleaved fragments of the protein were extracted with hexane and subjected to HPLC, resulting in purification of a single component of 2.3 kDa. Chemical analyses revealed that the purified fragment contains the tetradecapeptide Lys-Ile-Tyr-Pro-Ala-Val-Glu-Gly-Arg-Ile-Lys-Phe-Ser, ethanolamine, glucosamine, mannose, inositol, palmitic acid, and stearic acid. The peptide sequence determined is identified at positions 540-523 in the primary structure deduced from the cDNA sequence, which predicts a further extension to position 548, containing the hydrophobic amino acid sequence. Thus, it is concluded that the nmature 5’-nucleotidase lacks the predicted COOH-terminal peptide extension (524-548), which has been replaced by the glycophospholipid functioning as the membrane anchor of 5’-nucleotidase. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00142956
- Volume :
- 191
- Issue :
- 3
- Database :
- Academic Search Index
- Journal :
- European Journal of Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 13748801
- Full Text :
- https://doi.org/10.1111/j.1432-1033.1990.tb19158.x