7q02
Zn-free structure of lipocalin-like Milk protein, inspired from Diploptera punctata, expressed in Saccharomyces cerevisiae
Structural highlights
FunctionPublication Abstract from PubMedBACKGROUND: The Pacific Beetle Cockroach is the only known viviparous cockroach. The pregnant females provide nutrition to the embryos by secreting milk proteins (Lili-Mips), which crystallize in vivo. The crystals that grow in the embryo are heterogeneous in their protein sequence. It is not apparent from the structure determined what role heterogeneity and glycosylation played in crystallization. Lili-Mips are very nutritious. METHODS: Here, we report the cloning of synthesized Lili-Mip genes, their expression in Saccharomyces cerevisiae as secreted proteins, purification, crystallization, and the determination of a three-dimensional structure of one glycosylated and one deglycosylated form. RESULTS: A 2.35 A structure of the glycosylated form is bound to palmitoleic acid and has several Zn atom mediated interactions. A 1.45 A structure of the deglycosylated protein reveals a binding pocket that has both oleic and palmitoleic acid bound. Mass-spectrometry shows that oleic acid and palmitoleic acid are bound to the protein. Docking studies suggest that aliphatic chains of lengths 15, 16, and 18 carbons bind well in the pocket. CONCLUSIONS: The recombinantly expressed and secreted protein is glycosylated, has a bound fatty acid, is homogenous in its protein sequences, and readily forms crystals. The deglycosylated protein also crystallizes readily, suggesting that the high crystallizability of this protein is independent of glycosylation. GENERAL SIGNIFICANCE: Lili-Mips belong to the ubiquitous lipocalin family of proteins that bind to a large variety of ligands. While the residues lining the barrel are essential for the affinity of the ligand, our results show the role of side-chain orientations to ligand selectivity. Structure of recombinantly expressed cockroach Lili-Mip protein in glycosylated and deglycosylated forms.,KanagaVijayan D, Subramanian R, Santhakumari PR, Chavas LMG, Subramanian R, Banerjee S Biochim Biophys Acta Gen Subj. 2022 Mar;1866(3):130064. doi:, 10.1016/j.bbagen.2021.130064. Epub 2021 Dec 24. PMID:34958847[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
| ||||||||||||||||||||
