Lili-Mip:Milk Protein
BackgroundBackground
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The milk protein owes its origins to the viviparity of the Pacific beetle cockroach Diploptera punctate.[1] This species gives birth to live young and utilizes a reproductive strategy, viviparity, in which a supply of nutrients is provided to growing embryos in gestation periods of their lives. Lili Mip analysis revealed that milk proteins contain all the essential amino acids which is a vital feature.[2]
The time to reproductive maturity is significantly reduced because of viviparity. This enhancement of larval development provides a 60-fold whole body increase in protein during embryonic development. The viviparity of this cockroach species involves the rapid development of embryos that are capable of drinking and storing complex nutrients concentrated in crystalline form.[1]
While few studies has been conducted on the Lili-Mip, the direct crystallographic phasing and structure determination from crystals naturally grown in vivo rather than in vitro from over expressed proteins is the first report of its kind.[1]
Structure of Lili MipStructure of Lili Mip
Twenty-five distinct Lili-Mip complementary DNAs have been cloned and partially characterized that encode twenty-two Milk proteins. These peptides are each 171 amino acids long.[1]
Lili-Mip has been characterized as part of the lipocalin family consisting of one and to coordinate the lipid. The lipocalin family typically consists of lipophilic ligands in a cavity shaped by a common fold composed of a central Beta barrel.[2] The significant differences between lipocalin-like proteins is the hydrophobic pocket used for lipid coordination and categorizes the type of ligand that can be accommodated. Lipocalin proteins are known to carry fatty acids and other lipophilic materials.[3][4]
Electron density maps of the structure revealed densities for glycosylation at Asn35, Asn66, Asn79 and Asn145. N-acetyl-D-glucosamine were found to be unique ligands associated with Lili-Mip. One Beta-mannose (BMA) and two NAGS were identified at positions 35 and 79. Several other NAGs were identified at Asn145 (two) and one at Asn66.2 Models also coordinate lipids or inside the hydrophobic pocket of Lili Mip. The pocket in the Lili-Mip structure is 15 Angstrom’s deep and can accommodate up to 18-carbon fatty acid chain ligands. , notably through the formation of a stable π-stacking interaction of Tyr88 and Phe100 rings that restrict the length of the lipid.[2]
Milk proteins are rich in leucine, valine, asparagine, lysine, proline, and tyrosine residues, each accounting for 7-8% of the amino acids. However, they are lower in methionine and tryptophan residues being only about 0.5% each.[1] Lipids make up 16-22% of the dry weight, with cholesterol being the only steroid and linoleic acid the most abundant fatty acid. Fatty-acid chains in the barrel of Lili-Mip is presumed to be either linoleic acid or oleic acid.[5]
FunctionFunction
Lili-Mip is a complete nutritious material as it provides all of the essential amino acids, carbohydrates from the attached glycans, and essential lipids. Lili-Mips are the major nutrient source for growth for embryos.
While measuring nutritional value based on the presence of essential amino acids, there are eight conserved regions in Lili-Mip which suggest the Milk proteins might have additional functionality.[1]
The abundance of linoleic acid in the hydrophobic pocket of milk protein Lili-Mip of pregnant D. punctata is significant as linoleic acid is essential to most insects and other animals lipocalins may transport hydrophobic molecules such as cholesterol and linoleic acid that cannot be synthesized by insects.[2]
The relationship between lipocalin family of proteins and Lili Mip may suggest that Lili Mip may function in the transport of cholesterol to the embryos as lipocalin proteins have been documented to bind a variety of hydrophobic molecules including cholesterol.[2]
Heterogeneity in Lili-Mip amino-acid sequenceHeterogeneity in Lili-Mip amino-acid sequence
Heterogeneity is observed in the primary protein structure evident by glycosylation and lipid content of Lili-Mip; however, the function of heterogeneity is currently unclear.[2] Lili-Mip crystals containing multiple proteins with differing primary amino-acid sequences have been identified, heterogeneity for six of the 28 residues could be visualized clearly in the electron density maps.[2]
The significance of heterogeneity here is that there is no identified or reported crystal structure that has heterogeneity in protein sequence, carbohydrate, and lipid content together.[2] There are some examples of heterogeneity in protein sequence but with glycosylation and ligand binding independently. Also notable is the ability for Lili-Mip’s ability to diffract to atomic resolution.[2] The molecular weight ranges from 27 to 37 kDa which can be partially explained by the differential glycosylation of Milk proteins. The Lili-Mip contain four to six potential glycosylation sites that might be differentially glycosylated depending on the protein folding.[1] Furthermore, all heterogeneous residues of the protein are located on the surface of and not inside the barrel. This suggests the binding pocket is conserved.[2]
Medical ImplicationsMedical Implications
According to the UNICEF, nearly half of all deaths in children under the age of five are attributable to under nutrition. This organization and the World Health Organization’s data suggests this translates to about three-million deaths of children each year. Strategies to treat this malnutrition are vital to the survival of millions of people. Given the discovery of the nutrient content of Lili-Mip has opened doors into research about reproducibility to treat malnutrition and other protein-deficiency diseases. While milking cockroaches may not be the most productive, the discovery of this Milk protein may pave the way for an efficient method to reproducing it in mass. Cockroaches have been used in Brazil to make flour and resulted in far higher protein content than commercial flour from wheat (63.22% vs 9.8%). The cockroach flour contains eight essential amino acids and omega-3 and omega-9 fatty acids. Unsaturated fats were also found in this superfood like palmitic acid, oleic acid, and some saturated fats, all are considered parts of healthy diets.[6]
Further inquiry into the health implications of cockroaches and its milk should be conducted along with how to incorporate the nutrient rich food into many different diets.
Environmental ImplicationsEnvironmental Implications
Dairy milk and other alternatives are currently under environmental scrutiny as the amount of greenhouse gases produced by livestock raised for their milk and meat is too significant to ignore. While cockroach Diploptera punctate is far from the planet’s only solution to alleviate the stress of the accumulation of greenhouse gasses, there is potential for further research of the Milk protein produced by D. punctate as a more sustainable nutritious substances for our diets.
3D structures of milk protein3D structures of milk protein
Updated on 18-June-2023
4nyq, 4nyr, 5epq, 7bkx, 7qo2 – Mip – pacific beetle cockroach
ReferencesReferences
- ↑ 1.0 1.1 1.2 1.3 1.4 1.5 1.6 Banerjee S, Coussens NP, Gallat FX, Sathyanarayanan N, Srikanth J, Yagi KJ, Gray JS, Tobe SS, Stay B, Chavas LM, Ramaswamy S. Structure of a heterogeneous, glycosylated, lipid-bound, in vivo-grown protein crystal at atomic resolution from the viviparous cockroach Diploptera punctata. IUCrJ. 2016 Jun 27;3(Pt 4):282-93. doi: 10.1107/S2052252516008903. eCollection, 2016 Jul 1. PMID:27437115 doi:http://dx.doi.org/10.1107/S2052252516008903
- ↑ 2.00 2.01 2.02 2.03 2.04 2.05 2.06 2.07 2.08 2.09 Williford A, Stay B, Bhattacharya D. Evolution of a novel function: nutritive milk in the viviparous cockroach, Diploptera punctata. Evol Dev. 2004 Mar-Apr;6(2):67-77. PMID:15009119
- ↑ Salier JP, Akerstrom B, Borregaard N, Flower DR. Lipocalins in bioscience: the first family gathering. Bioessays. 2004 Apr;26(4):456-8. doi: 10.1002/bies.20013. PMID:15057944 doi:http://dx.doi.org/10.1002/bies.20013
- ↑ Flower DR, North AC, Attwood TK. Structure and sequence relationships in the lipocalins and related proteins. Protein Sci. 1993 May;2(5):753-61. PMID:7684291 doi:http://dx.doi.org/10.1002/pro.5560020507
- ↑ doi:10.1016/0020-1790(77)90023-3
- ↑ doi:10.1016/j.ifset.2017.08.015