Cloning, overexpression, purification, crystallization and preliminary X-ray analysis of a female-specific lipocalin (FLP) expressed in the lacrimal glands of Syrian hamsters

被引：1

作者：

Dubey, Ved Prakash ^{[1
]}

Pal, Biswajit ^{[1
]}

Srikantan, Subramanya ^{[1
]}

Pottabathini, Sambhavi ^{[1
]}

De, Prabir Kumar ^{[1
]}

Sankaranarayanan, Rajan ^{[1
]}

机构：

[1] Ctr Cellular & Mol Biol, Council Sci & Ind Res, Hyderabad 500007, Andhra Pradesh, India

来源：

ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | 2010年 / 66卷

关键词：

MALE-SPECIFIC PROTEINS; SUBMANDIBULAR-GLAND; CRYSTAL-STRUCTURE; BINDING-PROTEINS; CDNA CLONING; APHRODISIN; REPRESSION;

D O I：

10.1107/S1744309110008237

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Proteins belonging to the lipocalin superfamily are usually secretory proteins of molecular mass similar to 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 angstrom resolution. Solvent-content analysis indicated the presence of one monomer in the asymmetric unit.

引用

页码：509 / 512

页数：4

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