Impaired secretion of the elongated mutant of protein C (protein C-nagoya)

Molecular and cellular basis for hereditary protein C deficiency

Koji Yamamoto, Mitsune Tanimoto, Nobuhiko Emi, Tadashi Matsushita, Junki Takamatsu, Hidehiko Saito

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Genetic analysis of a heterozygous protein C-deficient patient revealed a novel deletion of a single guanine residue (8857G) among four consecutive guanine nucleotides [380TrP(TGG)-381Gly(GGT)] in exon IX, which encodes the carboxyl-terminal region of protein C. This deletion results in a frameshift mutation and substitution of the last 39 amino acids (381Gly-419Pro) with 81 abnormal amino acid residues, and we have designated this elongated variant as Protein C-Nagoya. A mutagenic primer was designed which replaced the third guanine residue upstream from the deletion with cytosine, thereby creating a new Aval site in an otherwise normal allele. Analysis of the polymerase chain reaction products derived from this, mutagenic primer showed that the abnormal allele has been inherited in this family. To elucidate how this molecular abnormality leads to protein C deficiency, an expression plasmid containing this mutation was transfected into COS 7, BHK, and ψ-2 cells, and the secretory process of the expressed Protein C-Nagoya was analyzed. ELISA and immunoprecipitation analysis with [35S]methionine labeling indicated that the mutant protein C, which was larger in size than normal, was mostly retained within the cells, and only a small portion of it was secreted into the medium. These results suggest that most of Protein C-Nagoya undergoes degradation within the producing cells, and this frameshift mutation apparently leads to protein C deficiency by impairment of secretion of the elongated protein C into plasma.

Original languageEnglish
Pages (from-to)2439-2446
Number of pages8
JournalJournal of Clinical Investigation
Volume90
Issue number6
Publication statusPublished - Dec 1992
Externally publishedYes

Fingerprint

Protein C Deficiency
Mutant Proteins
Protein C
Frameshift Mutation
Guanine
Alleles
Amino Acids
Guanine Nucleotides
Secretory Pathway
Cytosine
Immunoprecipitation
Methionine
Exons
Plasmids
Enzyme-Linked Immunosorbent Assay
Polymerase Chain Reaction
Mutation

Keywords

  • Expression study
  • Family diagnosis
  • Frameshift mutation
  • In vitro mutagenesis
  • Thrombophilia

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Impaired secretion of the elongated mutant of protein C (protein C-nagoya) : Molecular and cellular basis for hereditary protein C deficiency. / Yamamoto, Koji; Tanimoto, Mitsune; Emi, Nobuhiko; Matsushita, Tadashi; Takamatsu, Junki; Saito, Hidehiko.

In: Journal of Clinical Investigation, Vol. 90, No. 6, 12.1992, p. 2439-2446.

Research output: Contribution to journalArticle

Yamamoto, Koji ; Tanimoto, Mitsune ; Emi, Nobuhiko ; Matsushita, Tadashi ; Takamatsu, Junki ; Saito, Hidehiko. / Impaired secretion of the elongated mutant of protein C (protein C-nagoya) : Molecular and cellular basis for hereditary protein C deficiency. In: Journal of Clinical Investigation. 1992 ; Vol. 90, No. 6. pp. 2439-2446.
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