title: Characterization of the oligomerization and ligand-binding properties of recombinant rat lipocalin 11.
abstract: Lipocalin 11 (Lcn11), a recently identified member of the lipocalin family, potentially plays crucial physiological roles in male reproduction. In this present work, we cloned, expressed and purified the rat Lcn11 (rLcn11) protein Escherichia coli. A C59A/C156A substitution was introduced to ameliorate the misfolding and aggregation problem associated with the wild-type protein. From circular dichroism and non-reducing SDS-PAGE, we characterized the conformational properties of rLcn11 as a typical lipocalin scaffold with the conserved disulfide bridge. The results obtained from size-exclusion chromatography, cross-linking experiment and dynamic light scattering analysis indicate that the recombinant rLcn11 protein forms dimer in neutral solution. By using fluorescent probe-anilino-1 napthahlene sulfonic acid (ANS), we found rLcn might contain multiple hydrophobic binding sites for ligand binding. Similarly to the odorant-binding protein, rLcn11 processes a moderate affinity for binding 1-aminoanthracene (AMA), implying that Lcn11 might work as a dimeric chemoreception protein in male reproductive.
authors: Yan
authors: Y
authors: Gu
authors: Qiang
authors: Q
authors: Liu
authors: Peiyan
authors: P
authors: Chen
authors: Chenyun
authors: C
authors: Guo
authors: Yan
authors: Y
authors: Liu
authors: Yufen
authors: Y
authors: Zhao
authors: Yonglian
authors: Y
authors: Zhang
authors: Donghai
authors: D
authors: Lin
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D000873:Anthracenes / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D054834:Lipocalins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008297:Male; D020125:Mutation, Missense; D055503:Protein Multimerization / Q000502:physiology*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012098:Reproduction / Q000502:physiology
chemical_list: D000873:Anthracenes; D005456:Fluorescent Dyes; D054834:Lipocalins; D011994:Recombinant Proteins; C049062:1-aminoanthracene