Bottom-Up Proteomics Approach to Identify Fish Proteins in Tenualosa Ilisha and Salmo Salar Linnaeus
Disciplines
Biochemistry | Food Chemistry
Abstract (300 words maximum)
Proteomics investigation assists to identify and quantify all proteins available in a cell, tissue, or in an organism. Fish is an excellent source of protein and plays a critical role in global food security. Although proteomics studies have been widely used for human diseases and biomedical applications, only few research was reported on fish proteomics. In this study, we employed bottom-up proteomics approach to identify the key proteins present in Ilisha and Atlantic Salmon, two popular fishes found in Indian and Atlantic Oceans, respectively. These fishes live in the ocean but they move to freshwater rivers or lakes to spawn. To extract proteins, the fish tissue samples were disrupted using a Lysis buffer, then subsequently reduced, alkylated, and digested with trypsin/Lys-C protease mix. The peptides were separated by RP-LC using Vanquish Flex HPLC with a 90 min gradient. An Orbitrap Exploris 240 Mass Spectrometer was used to identify the peptides employing data-dependent analysis method. Proteome Discovery Software was used to search the LC-MS/MS data against the Ilisha and Salmon proteomes from NCBI database using the SEQUEST algorithm. Our preliminary results identified 37 proteins out of 103 in Ilisha and 52 proteins out of 151 in Salmon. The most prevalent protein in Ilisha was Lactate dehydrogenase (LDH) with a sequence coverage of 90%. LDH is a vital protein found in most tissues, that is responsible for converting sugar into cellular energy. On the other hand, Salmon’s highest coverage (59%) protein was Glyceraldehyde 3-phosphate. This protein is present during glycolysis and serves as an enzyme that catalyzes the breakdown of glucose.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad Halim
Bottom-Up Proteomics Approach to Identify Fish Proteins in Tenualosa Ilisha and Salmo Salar Linnaeus
Proteomics investigation assists to identify and quantify all proteins available in a cell, tissue, or in an organism. Fish is an excellent source of protein and plays a critical role in global food security. Although proteomics studies have been widely used for human diseases and biomedical applications, only few research was reported on fish proteomics. In this study, we employed bottom-up proteomics approach to identify the key proteins present in Ilisha and Atlantic Salmon, two popular fishes found in Indian and Atlantic Oceans, respectively. These fishes live in the ocean but they move to freshwater rivers or lakes to spawn. To extract proteins, the fish tissue samples were disrupted using a Lysis buffer, then subsequently reduced, alkylated, and digested with trypsin/Lys-C protease mix. The peptides were separated by RP-LC using Vanquish Flex HPLC with a 90 min gradient. An Orbitrap Exploris 240 Mass Spectrometer was used to identify the peptides employing data-dependent analysis method. Proteome Discovery Software was used to search the LC-MS/MS data against the Ilisha and Salmon proteomes from NCBI database using the SEQUEST algorithm. Our preliminary results identified 37 proteins out of 103 in Ilisha and 52 proteins out of 151 in Salmon. The most prevalent protein in Ilisha was Lactate dehydrogenase (LDH) with a sequence coverage of 90%. LDH is a vital protein found in most tissues, that is responsible for converting sugar into cellular energy. On the other hand, Salmon’s highest coverage (59%) protein was Glyceraldehyde 3-phosphate. This protein is present during glycolysis and serves as an enzyme that catalyzes the breakdown of glucose.