亲爱的研友该休息了!由于当前在线用户较少,发布求助请尽量完整地填写文献信息,科研通机器人24小时在线,伴您度过漫漫科研夜!身体可是革命的本钱,早点休息,好梦!

Application of mass spectrometry-based proteomics to muscle biology and meat science

肉的嫩度 肉类包装业 蛋白质组学 卡尔帕因 生物技术 质量(理念) 温柔 蛋白质降解 食品安全 牲畜 食品科学 生物 生化工程 生物化学 工程类 基因 生态学 哲学 认识论
作者
Logan Gregory Johnson
标识
DOI:10.31274/td-20240617-158
摘要

Livestock, meat, and food production systems have improved and become more efficient in the last few decades. The efficiency gains are partly due to the fundamental knowledge of the impacts of genetic selection, nutritional strategies, health interventions, facilities and welfare, and other technological innovations on the growth and efficiency of livestock. The eating quality of meat products remains challenging to predict, and undesirable meat products contribute to the overall waste of the production systems. Improving meat product quality could help reduce food product waste, improve access to meat products by a growing global population, and increase the value of meat products. Fundamental knowledge of factors impacting and relating to meat quality must be developed to balance productivity gains with value-related economic factors. A large body of research has documented that changes in the molecular features of postmortem muscle, including pH decline, lipid and protein oxidation, and protein degradation, result in measurable impacts on meat quality. Specifically, the degradation of structural proteins in postmortem muscle improves tenderness. Calpain proteinases contribute to postmortem proteolysis and degradation of specific cytoskeletal and myofibrillar proteins, which are related to improved meat quality and tenderness. Recognition of the molecular factors that can affect calpain activity and the susceptibility of proteins to degradation by calpains will generate robust information that can extend the knowledge of postmortem muscle biochemistry. In characterizing better these molecular factors, more robust and reliable tools to genetically select for meat quality and to categorize meat products based on their quality. The application and utility of mass spectrometry-based methods can allow for discovering and characterizing proteomic features associated with postmortem meat quality in a non-targeted or shot-gun approach. These mass spectrometry platforms can also be applied to hypothesis-based research questions. Both approaches have and will continue to provide crucial molecular information to improve the efficient production of livestock and meat products. This dissertation will explore both strategies of broader, discovery-based, and more targeted approaches to provide a greater understanding of proteome and protein differences related to postmortem muscle and meat quality. Therefore, the specific objectives of this dissertation were to 1) Identify the sarcoplasmic sub-proteomic differences between groups of aged pork chops based on water-holding capacity, 2) Determine the myofibrillar sub-proteomic differences between groups of pork chops based on instrumental tenderness values, 3) Understand the impacts of secondary lipid oxidation products on calpain-2 activity and autolysis and determine the quantity and localization of modification sites, and 4) Train machine learning models with specific tryptic peptide abundances to predict the ~2-week aged pork chop instrumental tenderness value. Lipid and protein oxidation occurs in postmortem skeletal muscle, and the products of these reactions generally accumulate during the postmortem period. The oxidation products of lipids can be aldehyde, ketone, and other reactive products, which have been shown to modify and impact proteins' functionality. Previous work has demonstrated that some lipid oxidation products can covalently modify calpain-1, resulting in activity and autolysis differences. The current work presented similar observations and showed that calpain-2 activity and the progression of autolysis were impacted differently by malondialdehyde, 2-hexenal, and 4-hydroxynonenal. Specific modification sites were determined with LC-MS/MS, including distinct malondialdehyde and 2-hexenal modification sites. Intact protein mass analysis with MALDI-MS revealed that many modifications on calpain-2 catalytic and regulatory subunits are likely to exist. These modifications can potentially occur in living and postmortem skeletal muscle, impact the solubility, functionality, and cellular localization of calpain-2 and other proteins, and alter the development of meat quality. Proteomic analysis using liquid chromatography-mass spectrometry (LC-MS) and tandem mass tags (TMT) has only recently been applied to research questions related to meat science. Herein, the utility of the fractionation of proteins based on solubility in a low-ionic strength buffer to reveal specific molecular differences related to pork quality is demonstrated. Indeed, this approach revealed potential proteolytic degradation products, including titin, desmin, and filamin-C, accumulating in the sarcoplasmic sub-proteome and more abundant in pork chop groups with less purge loss. Proteins related to the sarcoplasmic reticulum, calcium-regulating, and chaperone proteins, including calsequestrin, calcium transport ATPases, and heat shock proteins, were generally more abundant in these samples with less purge loss. In another experiment, the aged myofibrillar sub-proteome was fractionated to evaluate specific proteomic differences related to the instrumental star probe of pork chops. A lower abundance of proteins related to the Z-line and metabolic-associated proteins, such as titin, desmin, nebulin, PDZ and LIM domain type proteins, and glycogen phosphorylase, was observed in the myofibrillar sub-proteome of more tender pork chops. These observations help refine the knowledge of proteomic differences and changes related to pork quality. Proteomics experiments utilizing bottom-up approaches commonly result in an emphasis on protein abundance differences. In fact, two experiments in this dissertation have reported on differences in protein abundance. However, postmortem muscle's molecular changes and phenotype result in a more complex proteomic profile. The modifications of proteins, including conjugation, denaturation, oxidation, and degradation, generate a different profile of proteins, including intact and degraded forms of many proteins. The recognition of the limitations of current bottom-up approaches to identify degradation products led to the idea that the abundance of tryptic peptides generated from proteins soluble in a low-ionic strength buffer may be more informative and helpful in predicting the ultimate meat quality. Using feature-selected peptides, different machine learning models were trained and assessed to predict the ultimate instrumental star probe value. Machine learning models trained using only physical quality measures resulted in models with low (< 0.2) R2 values. The feature-selected peptides produced better fitting models, with higher (~ 0.5) R2 and lower root mean squared error and mean absolute error when predicting the continuous instrumental star probe value. The feature-selected peptides important in the machine learning models were identified to be primarily structural proteins, which are likely products of proteolysis in the aged sarcoplasmic sub-proteome. These data demonstrate the utility of specific tryptic peptides to predict pork tenderness. This research demonstrates the importance of understanding factors that influence and are related to meat quality differences. These molecular features are relevant and important in better characterizing the changes occurring in postmortem skeletal muscle. This work investigated proteomic differences in different protein fractions, factors that may impact calpain-2 functionality, and the utility of specific peptide features to improve the prediction of meat tenderness. Specific post-translational modifications and the solubility changes of protein and peptide degradation products are essential aspects less appreciated in the literature and warrant greater investigation, characterization, and validation. Notably, the work presented herein also provides a framework for future work that can refine understanding by applying fractionation of samples before bottom-up techniques. Top-down mass spectrometry approaches will become more commonly utilized techniques with improved instrumentation, and the work with these approaches lays the groundwork for subsequent research investigations.

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
duan完成签到 ,获得积分10
30秒前
significant完成签到,获得积分10
51秒前
55秒前
科研小趴菜完成签到,获得积分10
1分钟前
FengyaoWang完成签到,获得积分10
1分钟前
赘婿应助科研通管家采纳,获得10
2分钟前
2分钟前
raining_way完成签到 ,获得积分10
2分钟前
2分钟前
wzgkeyantong发布了新的文献求助20
2分钟前
由天与完成签到,获得积分10
2分钟前
多情易蓉发布了新的文献求助10
2分钟前
小榕树完成签到,获得积分10
3分钟前
aletta发布了新的文献求助20
3分钟前
3分钟前
研友_VZG7GZ应助科研通管家采纳,获得10
4分钟前
aletta完成签到,获得积分10
4分钟前
小胡爱科研完成签到 ,获得积分10
4分钟前
33发布了新的文献求助10
5分钟前
5分钟前
uhne发布了新的文献求助10
5分钟前
Hello应助科研通管家采纳,获得10
5分钟前
6分钟前
Leedesweet完成签到 ,获得积分10
6分钟前
Owen应助阿米尔盼盼采纳,获得10
6分钟前
彩虹儿应助33采纳,获得10
6分钟前
uhne完成签到 ,获得积分10
7分钟前
量子星尘发布了新的文献求助10
7分钟前
7分钟前
天天快乐应助科研通管家采纳,获得10
7分钟前
shipengfei完成签到 ,获得积分10
8分钟前
凤迎雪飘完成签到,获得积分10
8分钟前
周周完成签到 ,获得积分10
8分钟前
9分钟前
9分钟前
SciGPT应助小七采纳,获得10
9分钟前
Michael完成签到,获得积分10
10分钟前
ph完成签到 ,获得积分10
10分钟前
33发布了新的文献求助10
10分钟前
英俊的铭应助粽子采纳,获得10
11分钟前
高分求助中
(应助此贴封号)【重要!!请各用户(尤其是新用户)详细阅读】【科研通的精品贴汇总】 10000
二维材料在应力作用下的力学行为和层间耦合特性研究 600
苯丙氨酸解氨酶的祖先序列重建及其催化性能 500
Schifanoia : notizie dell'istituto di studi rinascimentali di Ferrara : 66/67, 1/2, 2024 470
Effects of different anesthesia methods on bleeding and prognosis in endoscopic sinus surgery: a meta-analysis and systematic review of randomized controlled trials 400
Laboratory Animal Technician TRAINING MANUAL WORKBOOK 2012 edtion 400
Progress and Regression 400
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 内科学 生物化学 物理 计算机科学 纳米技术 遗传学 基因 复合材料 化学工程 物理化学 病理 催化作用 免疫学 量子力学
热门帖子
关注 科研通微信公众号,转发送积分 4844701
求助须知:如何正确求助?哪些是违规求助? 4144922
关于积分的说明 12833830
捐赠科研通 3891609
什么是DOI,文献DOI怎么找? 2139216
邀请新用户注册赠送积分活动 1159249
关于科研通互助平台的介绍 1059536