PMID: 8262907

 

    Legend: Gene, Sites

Title : The structure of recombinant human carboxy-terminal-truncated macrophage colony-stimulating factor derived from mammalian cells

Abstract :
  1. The structure of recombinant human carboxy-terminal-truncated macrophage colony-stimulating factor expressed in CHO cells was investigated
  2. The bioactive protein ([-32-153] M-CSF ), expressed from a nucleotide sequence that encoded a signal peptide of 32 amino acids and N-terminal amino acids numbers 1-153, was heterogeneous in terms of molecular mass, as analyzed by SDS-PAGE, because of the presence of N-linked sugar moieties
  3. The primary structure of the polypeptide was determined by sequence analysis and amino acid analysis of the fragments obtained from lysylendopeptidase digests of reduced and alkylated M-CSF , and from pepsin digests of the intact molecule
  4. A sugar chain was located only at Asn-122 of the two putative sites of N-glycosylation that were present per subunit
  5. The homodimeric structure appeared to have seven disulfide bonds, formed by inter- or intra-molecular linkages, since there were no free thiol groups in the molecule
  6. The assignment of disulfide bonds by sequence analysis using peptide fragments indicated the combinations of Cys7-Cys90, Cys48-Cys139, and Cys102-Cys146
  7. Gel-filtration analysis of Ser31 [-32-153] M-CSF , in which the remaining Cys31 was replaced by Ser and which was expressed in COS cells, suggested that the mutein existed as a monomer
  8. Our study shows that the disulfide-bond pairings of [-32-153] M-CSF that is expressed and post-translationally modified in mammalian cells are identical to those of Escherichia coli-derived [3-153] M-CSF with only one intermolecular disulfide bond, namely, Cys31-Cys31
Output (sent_index, trigger, protein, sugar, site):
  • 4. Asn-122, , -, A sugar chain, Asn-122
  • 4. N-glycosylation, , -, -, sites
  • 4. located, , -, A sugar chain, Asn-122
Output(Part-Of) (sent_index, protein, site):
*Output_Site_Fusion* (sent_index, protein, sugar, site):
  • 4. colony-stimulating factor, A sugar chain, Asn-122

 

 

Protein NCBI ID SENTENCE INDEX