Cloning, overexpression, purification, crystallization and preliminary X-ray analysis of a female-specific lipocalin (FLP) expressed in the lacrimal glands of Syrian hamsters.
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.
Dubey
Ved Prakash
VP
Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research, Uppal Road, Hyderabad 500 007, India.
Pal
Biswajit
B
Srikantan
Subramanya
S
Pottabathini
Sambhavi
S
De
Prabir Kumar
PK
Sankaranarayanan
Rajan
R
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
D054834:Lipocalins
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