Chicken Egg Shell Membrane Associated Proteins and Peptides.
Egg shells are poultry industry byproducts with potential for use in various biological and agricultural applications. We have been interested in the membranes underlying the calcareous shell as a feed supplement, which showed potential to improve immunity and performance of post hatch poultry. Therefore, to determine their protein and peptide profiles, we extracted the egg shell membranes (ESM) from fresh unfertilized eggs with methanol and guanidine hydrochloride (GdHCl) to obtain soluble proteins for analysis by mass spectrometry. The methanol extract was subjected to matrix-assisted laser desorption ionization (MALDI), electrospray ionization (ESI), high-performance reverse phase liquid chromatographic separation (HPLC), and tandem mass spectrometry (MS/MS) to determine its peptide and protein profiles. The GdHCl extract was subjected to ESI-HPLC-MS/MS following trypsin digestion of reduced/alkylated proteins. Nine proteins from the methanol extract and >275 proteins from the GdHCl extract were tentatively identified. The results suggested the presence of several abundant proteins from egg whites, such as ovoalbumin, ovotransferrin, and lysozyme as well as many others associated with antimicrobial, biomechanical, cytoskeletal organizational, cell signaling, and enzyme activities. Collagens, keratin, agrin, and laminin were some of the structural proteins present in the ESM. The methanol-soluble fraction contained several clusterin peptides and defensins, particularly, two isoforms of gallin. The ratios of the two isoforms of gallin differed between the membranes obtained from brown and white eggs. The high abundance of several antimicrobial, immunomodulatory, and other bioactive proteins in the ESM along with its potential to entrap various microbes and antigens may make it a suitable vehicle for oral immunization of post hatch poultry and improve their disease resistance.
Poultry Production & Product Safety Research Unit, Agricultural Research Service, USDA; ‡Department of Poultry Science; #State Wide Mass Spectrometry Facility; and ⊥Cell & Molecular Biology Program, University of Arkansas , Fayetteville, Arkansas 72701, United States.
Sarbjeet
S
Makkar
Poultry Production & Product Safety Research Unit, Agricultural Research Service, USDA; ‡Department of Poultry Science; #State Wide Mass Spectrometry Facility; and ⊥Cell & Molecular Biology Program, University of Arkansas , Fayetteville, Arkansas 72701, United States.
Rohana
R
Liyanage
Poultry Production & Product Safety Research Unit, Agricultural Research Service, USDA; ‡Department of Poultry Science; #State Wide Mass Spectrometry Facility; and ⊥Cell & Molecular Biology Program, University of Arkansas , Fayetteville, Arkansas 72701, United States.
Lakshmi
L
Kannan
Poultry Production & Product Safety Research Unit, Agricultural Research Service, USDA; ‡Department of Poultry Science; #State Wide Mass Spectrometry Facility; and ⊥Cell & Molecular Biology Program, University of Arkansas , Fayetteville, Arkansas 72701, United States.
Balamurugan
B
Packialakshmi
Poultry Production & Product Safety Research Unit, Agricultural Research Service, USDA; ‡Department of Poultry Science; #State Wide Mass Spectrometry Facility; and ⊥Cell & Molecular Biology Program, University of Arkansas , Fayetteville, Arkansas 72701, United States.
Jack O
JO
Lay
Poultry Production & Product Safety Research Unit, Agricultural Research Service, USDA; ‡Department of Poultry Science; #State Wide Mass Spectrometry Facility; and ⊥Cell & Molecular Biology Program, University of Arkansas , Fayetteville, Arkansas 72701, United States.
Narayan C
NC
Rath
D000818:Animals; D002645:Chickens; D004528:Egg Shell / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D015017:Yolk Sac / Q000737:chemistry*
D008565:Membrane Proteins; D010455:Peptides; D011506:Proteins
egg shell; mass spectrometry; membrane; proteins