Lipocalins as a scaffold

Biochim Biophys Acta. 2000 Oct 18;1482(1-2):337-50. doi: 10.1016/s0167-4838(00)00145-x.

Abstract

The concept of scaffolds that can be equipped with artificial biochemically active sites has gained recent interest in the field of protein design. Members of the lipocalin protein family represent promising model systems in this respect. Especially prototypic lipocalins, such as the retinol-binding protein or the bilin-binding protein (BBP), exhibit a structurally simple one-domain fold with a conformationally well conserved beta-barrel as their central motif. This type of supersecondary structure is made of a cylindrically closed beta-sheet of eight antiparallel strands. At the open end of the barrel the beta-strands are connected by four loops in a pairwise manner so that a pocket for the ligand is formed. In a rational protein design study a metal-binding site was functionally grafted on the solvent-exposed surface of the beta-barrel, whereby the rigid backbone conformation permitted the spatially defined arrangement of three His side chains. In a combinatorial protein design approach, the natural ligand pocket of a lipocalin was reshaped. In this manner variants of the BBP were engineered which exhibit high affinity and remarkable specificity for haptens like fluorescein and digoxigenin. The so-called 'anticalins', i.e. artificial lipocalins recognizing prescribed ligands, could provide an interesting alternative to recombinant antibody fragments. Consequently, the use of lipocalins as a scaffold opens new applications for members of this functionally diverse protein family in biotechnology and medicine.

Publication types

  • Review

MeSH terms

  • Animals
  • Binding Sites
  • Carrier Proteins / chemistry*
  • Humans
  • Insect Proteins*
  • Ligands
  • Metals / metabolism
  • Models, Molecular
  • Protein Engineering
  • Protein Structure, Secondary
  • Retinol-Binding Proteins / chemistry*

Substances

  • Carrier Proteins
  • Insect Proteins
  • Ligands
  • Metals
  • Retinol-Binding Proteins
  • bilin-binding protein, insect