title: Conformational and biochemical characterization of a rat epididymis-specific lipocalin 12 expressed in Escherichia coli.
abstract: Lipocalin 12 (Lcn12) is a recently identified epididymis-specific protein that might play a significant physiological role in male reproduction. However, the detailed structure and function of Lcn12 remain to be determined. In the present work, we cloned, expressed, and purified the rat Lcn12 (rLcn12) protein in Escherichia coli, introduced the Cys176Ala substitution to eliminate the aggregation problem associated with the wild-type protein. Homology modeling results demonstrated that rLcn12 adopted an eight-stranded, antiparallel β-barrel conformation containing a conserved disulfide bond between Cys98 and Cys203, which was in accordance with the physicochemical properties elucidated by a combination of mass, circular dichroism, and nuclear magnetic resonance spectrometry. The purified rLcn12 protein exhibited a high binding affinity for all-trans retinoic acid in fluorescence titration experiments, implying that rLcn12 could be involved in retinoic acid transport in the epididymis.
authors.affiliation: Laboratory of Biomolecular NMR, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Peng
authors.forename: Jiafu
authors.initials: J
authors.lastname: Liu
authors.forename: Qiang
authors.initials: Q
authors.lastname: Liu
authors.forename: Yihe
authors.initials: Y
authors.lastname: Yao
authors.forename: Chenyun
authors.initials: C
authors.lastname: Guo
authors.forename: Yonglian
authors.initials: Y
authors.lastname: Zhang
authors.forename: Donghai
authors.initials: D
authors.lastname: Lin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D004220:Disulfides; D004822:Epididymis / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D054834:Lipocalins / Q000737:chemistry* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008297:Male; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D051381:Rats; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014212:Tretinoin / Q000737:chemistry
chemical_list: D004220:Disulfides; D054834:Lipocalins; D014212:Tretinoin