Phage display reveals a novel interaction of human tear lipocalin and thioredoxin which is relevant for ligand binding

FEBS Lett. 1999 Oct 22;460(1):182-6. doi: 10.1016/s0014-5793(99)01331-9.

Abstract

Human tear lipocalin (TL) is an unusual member of the lipocalin protein family, since it is known to bind a large variety of lipophilic ligands in vivo and acts as a cysteine proteinase inhibitor in vitro. It is suggested to function as a physiological protection factor by scavenging lipophilic potentially harmful compounds. Since protein-protein interaction or macromolecular complexation is a common feature of many lipocalins, we applied phage display technology to identify TL interacting proteins. By panning of a human prostate cDNA phagemid library against purified TL we isolated a thioredoxin (Trx) encoding phage clone. Biochemical analysis revealed that TL indeed interacts with Trx and is reduced by this redox protein. Reduction of the TL-specific disulfide bond is of functional relevance, since the reduced protein shows a nine-fold increase in ligand affinity when tested with retinoic acid as ligand.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Carrier Proteins / chemistry*
  • Carrier Proteins / genetics
  • Cloning, Molecular
  • DNA, Complementary / isolation & purification
  • Disulfides / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Eye Proteins / chemistry
  • Fluorescence
  • Gene Library
  • Humans
  • Ligands
  • Lipocalin 1
  • Male
  • Molecular Sequence Data
  • Oxidation-Reduction
  • Protein Binding
  • Thioredoxins / chemistry*
  • Thioredoxins / genetics
  • Tretinoin / chemistry

Substances

  • Carrier Proteins
  • DNA, Complementary
  • Disulfides
  • Eye Proteins
  • LCN1 protein, human
  • Ligands
  • Lipocalin 1
  • tear proteins
  • Thioredoxins
  • Tretinoin