Comprehensive Proteomic Analysis of Tenualosa Ilisha Employing Bottom-Up Proteomics Approach
Disciplines
Biochemistry | Molecular Biology | Other Biochemistry, Biophysics, and Structural Biology
Abstract (300 words maximum)
Proteomics study is an essential approach for molecular characterization, enabling the identification and quantification of proteins in different organisms, including fish. Fish known for being a high-quality protein source, is crucial for ensuring global food safety. Despite proteomics studies being widely used in human diseases, fish proteomics gets less attention. In this study, we utilized a bottom-up proteomics approach to identify the essential proteins in Tenualosa ilisha, a species belonging to the Clupeidae family and popularly known as ilishi, hilsa, hilsa herring, or hilsa shad, which is mostly caught in rivers and coastal areas in south Asian countries. In this context, we collected the tissues from different parts of the hilsha fish including the egg, brain, and tail. Afterward, we extracted the protein from these parts using lysis buffer followed by treating them with trypsin/Lys-C protease mix after reduction, alkylation, and digestion. Peptides were subsequently separated using reverse-phase liquid chromatography (RP-LC) using a Vanquish Flex HPLC system with a 90-minute gradient. Peptide identification was conducted with an Orbitrap Exploris 240 Mass Spectrometer utilizing a data-dependent analytical approach. The Proteome Discoverer software (V2.5) was used to analyze LC-MS/MS data against all proteins of Tenualosa ilisha (732 proteins) in the NCBI database with the SEQUEST algorithm. Our Initial findings showed the detection of 77 proteins in the ilisha tail tissue sample, 71 in the egg sample, and 66 in the brain sample with high confidence. Whereas, 120 proteins were identified in the tissue sample with medium confidence, along with 115 in the egg sample and 113 in the brain sample. Adenylate Kinase was the most prevalent protein found in Tenulosa ilisha, with a sequence coverage of 100%. Adenylate kinase (AK) acts as a signaling protein that is essential for controlling the balance of energy within cells.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad A. Halim
Comprehensive Proteomic Analysis of Tenualosa Ilisha Employing Bottom-Up Proteomics Approach
Proteomics study is an essential approach for molecular characterization, enabling the identification and quantification of proteins in different organisms, including fish. Fish known for being a high-quality protein source, is crucial for ensuring global food safety. Despite proteomics studies being widely used in human diseases, fish proteomics gets less attention. In this study, we utilized a bottom-up proteomics approach to identify the essential proteins in Tenualosa ilisha, a species belonging to the Clupeidae family and popularly known as ilishi, hilsa, hilsa herring, or hilsa shad, which is mostly caught in rivers and coastal areas in south Asian countries. In this context, we collected the tissues from different parts of the hilsha fish including the egg, brain, and tail. Afterward, we extracted the protein from these parts using lysis buffer followed by treating them with trypsin/Lys-C protease mix after reduction, alkylation, and digestion. Peptides were subsequently separated using reverse-phase liquid chromatography (RP-LC) using a Vanquish Flex HPLC system with a 90-minute gradient. Peptide identification was conducted with an Orbitrap Exploris 240 Mass Spectrometer utilizing a data-dependent analytical approach. The Proteome Discoverer software (V2.5) was used to analyze LC-MS/MS data against all proteins of Tenualosa ilisha (732 proteins) in the NCBI database with the SEQUEST algorithm. Our Initial findings showed the detection of 77 proteins in the ilisha tail tissue sample, 71 in the egg sample, and 66 in the brain sample with high confidence. Whereas, 120 proteins were identified in the tissue sample with medium confidence, along with 115 in the egg sample and 113 in the brain sample. Adenylate Kinase was the most prevalent protein found in Tenulosa ilisha, with a sequence coverage of 100%. Adenylate kinase (AK) acts as a signaling protein that is essential for controlling the balance of energy within cells.