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cDNA sequence for human erythrocyte ankyrin.

S Lambert1, H Yu, J T Prchal

  • 1Department of Biomedical Research, Saint Elizabeth's Hospital of Boston, Tufts University Medical School, MA 02135.

Proceedings of the National Academy of Sciences of the United States of America
|March 1, 1990
PubMed
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Researchers isolated human erythrocyte ankyrin cDNA, revealing a 206 kDa protein with 194 matching amino acids. This ankyrin gene is located on chromosome 8.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Ankyrin is a crucial protein in red blood cell structure and function.
  • Understanding ankyrin's genetic basis is vital for comprehending cellular integrity.

Purpose of the Study:

  • To isolate and characterize the complementary DNA (cDNA) for human erythrocyte ankyrin.
  • To determine the full coding sequence and predict the protein's structure and mass.
  • To investigate ankyrin gene expression and chromosomal localization.

Main Methods:

  • Isolation of overlapping cDNA clones from a reticulocyte library.
  • DNA sequencing to determine the composite cDNA sequence and open reading frame.
  • Amino acid sequencing of proteolytic peptides for validation.

Related Experiment Videos

  • RNA blot analysis to detect ankyrin mRNA in different cell types.
  • Genomic DNA analysis using sorted human chromosomes for gene mapping.

    • Main Results:

    • A composite cDNA sequence of 5636 base pairs was obtained, encoding a 1879-amino acid polypeptide (206 kDa).
    • The deduced amino acid sequence showed 194 identical residues to experimentally determined peptides and contained 23 homologous repeat units in the band 3 binding domain.
    • Evidence of alternative mRNA processing was observed, leading to deletions affecting peptide size and the COOH terminus.
    • Ankyrin mRNA was detected in erythroid cells (with differential expression in early types) and human cerebellum.
    • The human ankyrin gene was mapped to chromosome 8.

    Conclusions:

    • The complete cDNA sequence for human erythrocyte ankyrin provides a detailed molecular blueprint.
    • Ankyrin exhibits structural complexity, including repeat units and evidence of mRNA processing.
    • Ankyrin expression is not limited to erythroid cells, and its gene is located on chromosome 8.