Current Proteomics - Volume 19, Issue 2, 2022

Proteomic tools were extensively used to understand the relationship between muscle proteome and conversion of muscle to meat, post-mortem proteolysis, meat texture, and variation in meat color. Developments in proteomic tools have also resulted in their application for addressing the safety and authenticity issues including meat species identification, detection of animal byproducts, non-meat ingredients and tissues in meat products, traceability, identification of genetically modified ingredients, chemical residues and other harmful substances. Proteomic tools are also being used in some of the potential areas like understanding the effect of animal transportation, stunning, slaughter stress, halal authentication and issues related to animal welfare. Emerging advances in proteomic and peptidomic technologies and their application in traceability, meat microbiology, safety and authentication are taking a major stride as an interesting and complementary alternative to DNA-based methods currently in use. Future research in meat science need to be linked to emerging metabolomic, lipidomic and other omic technologies for ensuring integrated meat quality and safety management. In this paper, a comprehensive overview of the use of proteomics for the assessment of quality and safety in the meat value chain and their potential application is discussed.

Background: Soil salinity is a major issue that seriously affects plant growth and cultivated land utilization. Salt tolerance is one of the most fundamental biological processes that ensures plant’s survival. SOS2 is one of the most important components of the Salt Overly Sensitive (SOS) signaling pathway, which maintains plant ion homeostasis under salt stress. The SOS2-related signaling pathways remain incompletely exploited especially at the proteomics level. Objective: In this paper, proteins potentially interacting with and regulated by SOS2 in Arabidopsis were identified. Methods: The proteomes of Arabidopsis Wild Type (WT) and SOS2-deficient mutant (sos2-2) exposed to 100 mM NaCl for 6 h were compared, proteins were identified using data-independent acquisition- based quantitative proteomics strategy. Results: A total of 7470 proteins were identified and quantified, 372 Differentially Expressed Proteins (DEP) were detected between WT and sos2-2 mutant under normal condition and 179 DEPs were identified under salt treatment. Functional analysis showed that the DEPs were mainly involved in protein binding and catalytic activity. Among the DEPs under salt stress, the protein expressions of AVP1, Photosystem II reaction center protein A, B, C, and stress-responsive protein (KIN2) were significantly up-regulated. LHCA1, LHCA2, LHCA4, ATPD and ATPE were significantly down-regulated. These proteins were involved in biological processes including: stress response, photosynthesis, transport and heat shock. Conclusion: These results revealed complexity of the functions of SOS2 in maintaining intracellular homeostasis, in addition to its function in sodium homeostasis. Plant salt resistance is not independent but closely related to metabolic processes including photosystem, ATP synthase, transport and other stress resistances.

Background: Heavy metals and metalloids like arsenic, cadmium, mercury acts as denaturing agent for biomolecules. They interfere with protein’s physiological activity by forming a complex with the protein’s side chain or removing the essential metal ions from metalloproteins and replacing them. Protein aggregation is an extensive phenomenon in a cell and is linked with various pathological conditions. Aim: In this study, we aim to prove that proteins are highly susceptible to arsenite toxicity by arsenite-induced protein aggregation; and that naringin reduces the aggregation effect. Methods: Several biophysical techniques were employed to study the protein aggregation due to arsenite and its prevention by naringin. Results: Through our experiments, the results showed that aggregation induced by arsenite was reduced in the presence of naringin at twice the concentration of arsenite. Conclusion: In conclusion, our study showed that naringin plays a protective role during HSA aggregation due to arsenite.

Aim: To evaluate the binding affinity and biological potency of gonadotropin-releasing hormone analogue (GnRHa) Buserelin (C60H86N16O13) based on in silico and in vivo testing for induced breeding in Clarias magur. Background: Many attempts have been made to induce C. magur, but encouraging results have not yet been achieved. Hence, it is the need of the hour to find out more potent analogues or other bio-molecules for induced breeding in C. magur to facilitate sustainable aquaculture. Objective: To determine the binding affinity of C. magur GnRH receptor through in silico and to validate it for induced breeding of C. magur. Methods: Buserelin (C60H86N16O13) was selected as the potential GnRHa after screening several peptides for their binding energy with the C. magur GnRH receptor. The induced breeding trial was set up at ICAR-CIFE Powarkheda Centre, M.P. India, and Buserelin was administered in different doses to the brooders along with the dopamine inhibitor domperidone. The standard treatment with the commercial salmon GnRH (sGnRH) analogue Ovaprim® (Syndel, USA) was used as the control. Results: The 3-D structure of C. magur GnRH receptor was generated using MODELLER software. Molecular docking studies revealed the binding preference of the receptor as chicken (c) Gn- RH-II > Buserelin > sGnRH > catfish (cf) GnRH > human (m) GnRH. Though Buserelin showed better binding affinity compared to sGnRH, induced breeding experiments with magur showed similar performance of the ligands at the equivalent dose of 20 μg/kg B.W., but the spontaneous release of milt from the males was not observed in both cases. Significantly better reproductive parameters were recorded with Buserelin at the dose of 30 μg/kg B.W. Conclusion: The study revealed that that the GnRHa Buserelin can be used as an effective inducing agent for breeding in C. magur.

Background: p16 is a tumor suppressor protein that is significantly involved in cycle regulation through the reduction of cell progression from the G1 phase to the S phase via CDK-cyclin D/p16^INK4a/pRb/E2F cascade. The minimum functional domain of p16 has been uncovered that may function comparable to wild type p16. Objective: To expand the knowledge on molecules and mechanisms by which p16 or p16^66-156 fragment suppresses human fibrosarcoma cell line growth, differential proteome profiles of fibrosarcoma cells following p16 full length or the functional domain overexpression, were analyzed. Methods: Following transfecting HT-1080 fibrosarcoma cells with p16 full length, p16^66-156 truncated form, and pcDNA3.1 empty vector, protein extract of each sample was harvested and clarified by centrifugation, and then the protein content was determined via Bradford assay. All protein extract of each sample was analyzed by two-dimensional gel electrophoresis. Immunoblot analysis was performed as further validation of the expression status of identified proteins. Results: Expression of p16 or p16^66-156 fragment could induce mostly the common alterations (up/- down-regulation) of proteome profile of HT-1080 cells. Mass spectrometry identification of the differentially expressed protein spots revealed several proteins that were grouped in functional clusters, including cell cycle regulation and proliferation, cell migration and structure, oxidative stress, protein metabolism, epigenetic regulation, and signal transduction. Conclusion: The minimum functional domain of p16 could act in the same way as p16 full length. Also, these new findings can significantly enrich the understanding of p16 growth-suppressive function at the molecular level by the introduction of potential candidate targets for new treatment strategies. Furthermore, the present study provides strong evidence on the functional efficacy of the identified fragment of p16 for further attempts toward peptidomimetic drug design or gene transfer to block cancer cell proliferation.

Background: The nasal fibroblast secretome, which includes various cytokines, chemokines, and growth factors, promotes cell migration. Currently, the proteomics of Airway Fibroblast (AF) Conditioned Medium (AFCM) are being actively studied. Objective: This study was aimed at profiling and identifying the AF secreted proteins that can enhance wound healing of the airway epithelium and predict the potential pathway involved. Methods: Airway Epithelial Cells (AECs) and AFs were isolated from redundant human nasal turbinate and cultured. AFCM was collected by culturing the AFs either with serum-free airway epithelium basal medium (AECM) or with serum-free F12:DMEM (FDCM). For evaluating cell migration, the AECs were supplemented with airway epithelium medium and defined keratinocyte medium (1:1; AEDK; control), or with AEDK supplemented with 20% AECM or 20% FDCM. The mass spectrometry sample was prepared by protein precipitation, followed by gel electrophoresis and in-gel digestion. Results: AECM promoted better cell migration compared to the FDCM and the control medium. Bioinformatics analysis identified a total of 121, and 92 proteins from AECM and FDCM, respectively: 109 and 82 were identified as secreted proteins, respectively. STRING® analysis predicted that 23 proteins from the AECM and 16 proteins from the FDCM are involved in wound healing. Conclusion: Conditioned medium promotes wound healing by enhancing cell migration, and we successfully identified various secretory proteins in a conditioned medium that play important roles in wound healing.