Proteome analysis of early post-mortem changes in two bovine muscle types:M. longissimus dorsi andM. semitendinosis

Abstract

To study early post-mortem changes in muscle tissues from bull calves, cytosole proteins from two muscles: M. longissimus dorsi (LD) and M. semitendinosis (ST) at 0 and 24 h after slaughter were analysed by 2-DE. Principal component analysis (PCA) and rotation testing were used to analyse the protein patterns in the two muscles in order to select protein spots that were significantly different at the two time-points. Selected proteins were identified by MALDI-TOF/TOF. Five proteins, namely cofilin, lactoylglutathione lyase, substrate protein of mitochondrial ATP-dependent proteinase SP-22, HSP 27 and HSP20, were changed in both LD and ST muscles during post-mortem storage. Fifteen additional protein changes were observed in either LD or ST muscles, and some of these changes have not previously been observed to change during post-mortem storage of bovine muscles. Further studies will reveal the relevance of these biomarkers for meat quality.

Heat Stabilization of the Tissue Proteome: A New Technology for Improved Proteomics

“…For example, the enkephalin neuropeptides (met-enk-RF and met-enk-RSL) were clearly detected in the stabilized samples and almost undetectable in the snap-frozen samples. Interestingly, the isotopically labeled neuropeptides added as internal standards were only detected in stabilized tissue, aside from the labeled protein fragment stathmin (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) which was detected independent of sample treatment, indicating ex vivo proteolytic activity and the relative stability of the stathmin (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) fragment.…”

“…Mice in a fourth sample group (n ) 4) were killed by focused microwave irradiation (1.4 s at 4.5-5 kW, Muromachi Kikai, Tokyo, Japan) which denature the proteins before dissection and homogenization. A microtip ultrasonicator (Vibra-Cell, Sonics & Materials) was used to homogenize the tissue in 5× the sample weight of 0.25% HAc extraction solution containing 40 mM of isotopically labeled stathmin (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20), met-enkephalin, neurotensin, substance P (1-11), and substance P (1-7) as internal standards. The crude extract was centrifuged at 20 000g for 30 min at 4°C to pellet cell debris and ultrafiltered using a 10 kDa centrifugal cutoff filter (Microcon YM-10, Millipore) to isolate the peptide content.…”

Skeletal muscle proteome and meat quality of broiler chickens subjected to gas stunning prior slaughter or slaughtered without stunning

“…Creatine kinase M-type is an enzyme found in sarcolemma and in sarcoplasmic reticulum of muscle cells where it is functionally involved in the calcium transport and ATPase activity (Rossi, Eppenberger, Volpe, Cotrufo, & Wallimann, 1990). It is involved in the transfer of high-energy phosphate between creatine and ATP molecule (Jia, Hollung, Therkildsen, Hildrum, & Bendixen, 2006) and it is found especially in the tissues that have high demand for energy such as skeletal muscle, brain and kidney. The higher expression of creatine kinase M-type observed in the gas-stunned broilers could possibly be associated with higher demand for energy during hypoxia.…”

Impact of Temperature Dependent Sampling Procedures in Proteomics and Peptidomics – A Characterization of the Liver and Pancreas Post Mortem Degradome

“…5 and 6). Muscle and brain are the most characterized tissues with regard to post mortem degradation, with most proteomics studies focused on brain tissue (1,(3)(4)(5)(11)(12)(13)(14)(15)(16). Several proteins found in the brain degradome were also detected in the liver or pancreas degradome (Table III, IV).…”

“…PMI degradation has mainly been studied on human or mouse brain tissue, using two-dimensional electrophoresis (2-DE), SDS-PAGE, and immunoblotting (1,(3)(4)(5)(6)(7)(8)(9)(10)(11)(12). There are also a few proteomic studies on muscle tissue degradation in livestock (13)(14)(15)(16).…”