Molecular cloning and primary structure of Kell blood group protein

Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6353-7. doi: 10.1073/pnas.88.14.6353.

Abstract

The Kell blood group is a major antigenic system in human erythrocytes. Kell antigens reside on a 93-kDa membrane glycoprotein that is surface-exposed and associated with the underlying cytoskeleton. We isolated tryptic peptides and, based on the amino acid sequence of one of the peptides and by using the PCR, prepared a specific oligonucleotide to screen a lambda gt10 human bone-marrow cDNA library. Four clones were isolated, one containing cDNA with an open reading frame for an 83-kDa protein. All known Kell amino acid sequences were present in the deduced sequence; moreover, rabbit antibody to a 30-amino acid peptide, prepared from this sequence, reacted on an immunoblot with authentic Kell protein. The Kell cDNA sequence predicts a 732-amino acid protein. Hydropathy analysis indicates a single membrane-spanning region, suggesting that Kell protein is oriented with 47 of its N-terminal amino acids in the cell cytoplasm, and a 665-amino acid segment, which contains six possible N-glycosylation sites, is located extracellularly. Computer-based search showed that Kell has structural and sequence homology to a family of zinc metalloglycoproteins with neutral endopeptidase activity.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal
  • Antigens, CD / genetics
  • Antigens, Differentiation / genetics
  • Antigens, Neoplasm / genetics
  • Base Sequence
  • Blotting, Northern
  • Bone Marrow / immunology
  • Cloning, Molecular
  • Gene Library
  • Humans
  • Kell Blood-Group System / genetics*
  • Molecular Sequence Data
  • Neprilysin
  • Oligonucleotide Probes
  • Polymerase Chain Reaction / methods
  • RNA / genetics
  • RNA / isolation & purification

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, Differentiation
  • Antigens, Neoplasm
  • Kell Blood-Group System
  • Oligonucleotide Probes
  • RNA
  • Neprilysin

Associated data

  • GENBANK/M64934