title: Cloning, overexpression, purification, crystallization and preliminary X-ray analysis of a female-specific lipocalin (FLP) expressed in the lacrimal glands of Syrian hamsters.
abstract: Proteins belonging to the lipocalin superfamily are usually secretory proteins of molecular mass approximately 20 kDa with a hydrophobic pocket for the binding and transport of diverse small ligands. Various lipocalins have been associated with many biological processes, e.g. immunomodulation, odorant transport, pheromonal activity, retinoid transport, cancer-cell interactions etc. However, the exact functions of many lipocalins and the ligands bound by them are unclear. Previously, the cDNA of a 20 kDa lipocalin (FLP) which is female-specifically expressed in the lacrimal glands of Syrian (golden) hamsters and secreted in the tears of females has been identified and cloned. His-tagged recombinant FLP (rFLP) has now been cloned, overexpressed in Escherichia coli as a soluble protein and purified to homogeneity using Ni-affinity followed by size-exclusion chromatography. Purified rFLP was crystallized using the sitting-drop vapour-diffusion method. The crystals tested belonged to space group P2(1)2(1)2(1) and diffracted to beyond 1.86 A resolution. Solvent-content analysis indicated the presence of one monomer in the asymmetric unit.
authors.affiliation: Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research, Uppal Road, Hyderabad 500 007, India.
authors.forename: Ved Prakash
authors.initials: VP
authors.lastname: Dubey
authors.forename: Biswajit
authors.initials: B
authors.lastname: Pal
authors.forename: Subramanya
authors.initials: S
authors.lastname: Srikantan
authors.forename: Sambhavi
authors.initials: S
authors.lastname: Pottabathini
authors.forename: Prabir Kumar
authors.initials: PK
authors.lastname: De
authors.forename: Rajan
authors.initials: R
authors.lastname: Sankaranarayanan
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D006224:Cricetinae; D003460:Crystallization; D018360:Crystallography, X-Ray; D005260:Female; D015870:Gene Expression; D007765:Lacrimal Apparatus / Q000737:chemistry*; D054834:Lipocalins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008647:Mesocricetus
chemical_list: D054834:Lipocalins
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