Characterization of the fertilized chicken egg yolk proteome and lipidome
In 2015, MYOS RENS researchers in collaboration with scientists at Rutgers University used a mass spectrometry-based proteomics approach to study the changes that take place in egg yolk during the first 24 hours following fertilization and published their findings in Food & Function. Fertilized egg yolk is used in the proprietary manufacturing process of Fortetropin®, the first clinically demonstrated, natural myostatin reducing agent.
The impact of fertilization on the chicken egg yolk plasma and granule proteome 24 hours post-lay at room temperature: capitalizing on high-pH/low-pH reverse phase chromatography in conjunction with tandem mass tag (TMT) technology
Neerav D. Padliya,*a Meiqian Qian,b Sushmita Mimi Roy,c Patrick Chu,d Haiyan Zheng,b Alex Tess,a Maghsoud Dariania and Robert J. Hariria
a Research & Development, MYOS RENS Technology Inc., Cedar Knolls, USA
b Biological Mass Spectrometry Facility, Rutgers, The State University of New Jersey, Piscataway, USA
c iON Bioservices, L.L.C., Sunnyvale, USA
d Sage-N Research, Inc., Milpitas, USA
Food Funct., 2015,6, 2303-2314
Received 23 Mar 2015, Accepted 08 Jun 2015
First published online 09 Jun 2015
Chicken egg yolk is a rich source of nutrients providing high quality proteins, vitamins, minerals, carotenoids and antioxidants. Chicken egg yolk, recovered from whole egg within 24 hours post-lay has been utilized as a starting material in the preparation of a dietary supplement that has been demonstrated to lead to gains in muscle mass in a human clinical study. Further, an oil derived from chicken egg yolk has been utilized as a topical agent to treat third degree burn injury. The molecular changes that take place in fertilized, chicken egg yolk during the first 24 hours post-lay are not well understood. By studying how the protein composition of egg yolk varies with fertility status, one can utilize this knowledge to develop egg yolk-based products that have been optimized for specific applications. In this study, a direct quantitative comparison was made between the proteome of fertilized chicken egg yolk and the proteome of unfertilized chicken egg yolk, both maintained at 20 °C and analyzed within 24 hours post-lay. Egg yolk proteins from each fertility state were digested with trypsin, labeled with distinct chemical labels (tandem mass tag reagents) and then combined in a 1:1 ratio. A TMT-labeled tryptic digest derived from chicken egg yolk proteins (fertilized and unfertilized) was separated using high-pH/low-pH reverse-phase chromatography and analyzed using mass spectrometry. 225 protein identifications were made from this TMT-labeled tryptic digest based on a minimum of 2 unique peptides observed per protein. 9 proteins increased in abundance in fertilized egg yolk relative to unfertilized egg yolk and 9 proteins decreased in abundance in fertilized egg yolk relative to unfertilized egg yolk. Some proteins that increased in abundance in fertilized egg yolk play an important role in angiogenesis (pleiotrophin, histidine rich glycoprotein) and defense against pathogens (mannose-binding lectin, β-defensin 11, serum amyloid P-component, ovostatin). Based on this study, fertilized chicken egg yolk may be more useful as a starting material relative to unfertilized chicken egg yolk for the purpose of enriching or isolating proteins with pro-angiogenic and anti-microbial properties.
Computational Design of Novel Peptide Inhibitors of Myostatin
Over the last several years, myostatin inhibition has emerged as a potential strategy for the treatment of cancer cachexia along with other muscle wasting disorders. Myostatin, a member of the TGF-β superfamily, inhibits the process of myogenesis, the development of muscle tissue. First-generation myostatin-directed monoclonal antibodies exhibited poor specificity towards myostatin with respect to other TGF-β ligands, giving rise to many undesired side effects such as the inhibition of wound healing. Rather than focus on antibodies, we have turned our attention towards the protein follistatin, a natural antagonist of myostatin which exists as three alternative splice variants, FS-288, FS-300 and FS-315 with the highest concentration found in the female ovaries followed by the skin. Specifically, we predicted peptides that the action of the proteases, pepsin followed by chymotrypsin on follistatin would give rise to and then modeled the binding of these follistatin-biomi metic peptides with myostatin. The secondary and tertiary structure of myostatin was modeled using the program, PHYRE2