Identification of a channel catfish, Ictalurus punctatus (Rafinesque), leukocyte-specific leucine zipper protein

Dev Comp Immunol. Mar-Apr 1999;23(2):149-63. doi: 10.1016/s0145-305x(98)00048-2.

Abstract

Five clones isolated from a channel catfish cDNA library were each reactive with monoclonal antibodies (mAbs) C3-1 and 51A only. The size of the cDNA inserts from C3-1 and 51A positive clones was 2.5 Kb and identical based on sequence analysis. Monoclonal antibodies C3-1 and 51A specifically reacted with the expressed product of the 2.5 Kb cDNA clone. The complete DNA sequence indicated that the 2.5 Kb cDNA encoded an approximately 50 Kd protein molecule consisting of 445 amino acids. Sequence analysis showed that this putative protein was a potential leucine-zipper DNA binding protein. Comparison of the deduced amino acid sequence demonstrated homology (14.6 to 19.5%) throughout the sequence of the catfish protein with a group of cytoplasmic-leucine zipper containing proteins of humans; paraneoplastic cellebellar degeneration related (cdr) antigen 2 and 3 with 39.8 to 56.3% homology in the leucine-zipper motif (amino acids 52 through 175 in the catfish protein). This protein was detected in nuclear extracts. cytoplasmic membrane preparations and cytosolic extracts of neutrophils and lymphocytes when reacted with mAbs C3-1 and 51A in an ELISA. However, the intensity of the reactions was dependent upon the cell type and cellular component. The putative cdr protein was not detected with any appreciable intensity in preparations from other cell types. This finding strongly suggests that this protein is expressed in a leukocyte-specific manner and is unique among the cdr group in that it is being expressed in a site that is not immune privileged.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • DNA, Complementary
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / immunology*
  • Gene Library
  • Ictaluridae / immunology*
  • Leucine Zippers*
  • Leukocytes / immunology*
  • Microscopy, Confocal
  • Molecular Sequence Data
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid

Substances

  • DNA, Complementary
  • DNA-Binding Proteins