Homology Modeling Study of Bovine μ-Calpain Inhibitor-Binding Domains

The activated mammalian CAPN-structures, the CAPN/CAST complex in particular, have become an invaluable target model using the structure-based virtual screening of drug candidates from the discovery phase to development for over-activated CAPN linked to several diseases, such as post-ischemic injury and cataract formation. The effect of Ca²⁺-binding to the enzyme is thought to include activation, as well as the dissociation, aggregation, and autolysis of small regular subunits. Unfortunately, the Ca²⁺-activated enzyme tends to aggregate when provided as a divalent ion at the high-concentration required for the protease crystallization. This is also makes it very difficult to crystallize the whole-length enzyme itself, as well as the enzyme-inhibitor complex. Several parameters that influence CAPN activity have been investigated to determine its roles in Ca²⁺-modulation, autoproteolysis, phosphorylation, and intracellular distribution and inhibition by its endogenous inhibitor CAST. CAST binds and inhibits CAPN via its CAPN-inhibitor domains (four repeating domains 1–4; CAST1–4) when CAPN is activated by Ca²⁺-binding. An important key to understanding CAPN1 inhibition by CAST is to determine how CAST interacts at the molecular level with CAPN1 to inhibit its protease activity. In this study, a 3D structure model of a CAPN1 bound bovine CAST4 complex was built by comparative modeling based on the only known template structure of a rat CAPN2/CAST4 complex. The complex model suggests certain residues of bovine CAST4, notably, the TIPPKYQ motif sequence, and the structural elements of these residues, which are important for CAPN1 inhibition. In particular, as CAST4 docks near the flexible active site of CAPN1, conformational changes at the interaction site after binding could be directly related to CAST4 inhibitory activity. These functional interfaces can serve as a guide to the site-mutagenesis in research on bovine CAPN1 structure-function relationships for the design of small molecules inhibitors to prevent uncontrolled and unspecific degradation in the proteolysis of key protease substrates.     

The comparison of three β-agonists for growth performance,carcass characteristics and meat quality of feedlot cattle

Forty-eight Bonsmara steers were assigned to three treatment groups and one control group consisting of 12 animals each. The control (C) received no β-agonist, while the three treatment groups received zilpaterol (6 ppm) (Z), ractopamine (30 ppm) (R) or clenbuterol (2 ppm) (Cl) for the last thirty days on feed. Growth performance (final 30 days), USDA quality and yield grades and meat quality (shear force, chemical, histological and biochemical) were compared for the three β-agonist and control groups. Animals responded negatively to Cl treatment during initial stages of supplementation, which was evident in lower feed consumption and initial growth rates. For carcass growth and yield, Cl had greater and more efficient growth rates, higher dressed out yields (proportional), lower USDA yield grades, and reduced marbling compared with C (P < 0.05). For meat quality measurements, the M. longissimus (LL) and M. semitendinosus (ST) were sampled. Cl had the greatest effect (P < 0.05) on WBSF, especially on the LL, followed by Z. Variation in tenderness and ageing effects corresponded with variation in calpastatin activity and myofibrillar fragmentation between treatment groups. While zilpaterol and ractopamine are currently the only products registered for cattle in different countries, it seems that zilpaterol has an advantage in carcass growth efficiency and yield without showing any adaptation problems for animals such as experienced by the more aggressive β-agonist clenbuterol.     

Effect of the insulin-like growth factor-II and RYR1 genotype in pigs on carcass and meat quality traits

Recently, a new QTN (quantitative trait nucleotide), which is located in the regulatory sequence of the imprinted IGF-II gene was discovered in the pig and is associated with a significant increase in IGF-II mRNA expression in skeletal muscle during postnatal growth. The aim of the current study was to investigate the effect of the IGF-II paternal allele (Apat and Gpat animals that inherited, respectively, the mutant and wild type paternal allele of interest) on carcass and meat quality traits in Nn and NN RYR1 genotypes. A total of 141 animals were measured, almost equally distributed over the IGF-II and RYR1 genotypes and gender. The Apat allele increased carcass lean meat percentage with approximately 4.5% (P < 0.001) as a result of decreased backfat thickness. Average live weight daily gain was not affected, hence average daily lean meat gain was significantly higher for Apat compared to Gpat animals. The IGF-II mutation had no noticeable effect on meat quality in contrast with the RYR1 mutation. No interaction effects of both mutations on meat quality were noticed.     

Variation in exon 10 of the ovine calpain 3 gene (CAPN3) and its association with meat yield in New Zealand Romney sheep

Variation in the ovine CAPN3 gene was analysed using PCR-single strand conformational polymorphism, and its effect on growth and carcass traits was assessed in 513 New Zealand Romney lambs produced by 17 unrelated rams. Among the four allelic variants detected, the presence of variant *02 was found to be associated with an increased proportion of shoulder yield (absent: 32.6 ± 0.01%; present: 33.4 ± 0.03%; P = 0.016), and tended to be associated with increased shoulder yield (lean meat yield of the shoulder expressed as a percentage of the hot carcass weight) (absent: 16.6 ± 0.06%; present: 17.02 ± 0.20%; P = 0.067). No association was detected with growth traits or other carcass traits.     

Protein Oxidation: Basic Principles and Implications for Meat Quality

The involvement of oxidized proteins to the development of biological diseases has been studied for a few decades, but the effects and the mechanisms of protein oxidation in food systems are largely unknown. Protein oxidation is defined as the covalent modification of a protein induced either by the direct reactions with reactive oxygen species (ROS) or indirect reactions with secondary by-products of oxidative stress. ROS can cause oxidation in both amino acid side chains and protein backbones, resulting in protein fragmentation or protein–protein cross-linkages. Although all amino acids can be modified by ROS, cysteine, and methionine that are the most susceptible to oxidative changes due to high reaction susceptibility of the sulfur group in those amino acids. Oxidative modifications of proteins can change their physical and chemical properties, including conformation, structure, solubility, susceptibility to proteolysis, and enzyme activities. These modifications can be involved in the regulation of fresh meat quality and influence the processing properties of meat products. Oxidative stress occurs when the formation of oxidants exceeds the ability of antioxidant systems to remove the ROS in organisms. Increased levels of protein oxidation have been associated with various biological consequences, including diseases and aging, in humans and other animal species. The basic principles and products of protein oxidation and the implications of protein oxidation in food systems, especially in meat, are discussed in this review.     

Biochemical and sensorial evaluation of intact and boned broiler breast meat tenderness during ageing

Meat tenderness is the main characteristic demanded by consumers and is affected by rigor mortis development and proteolysis activities, both of which occur during carcass refrigeration. In this work, we demonstrate that broiler breast fillet tenderness can be further increased and its extension depends on whether or not meat is excised from the carcass. Post-harvest samples taken from 0 to 72 h after slaughtering and kept refrigerated at 2 ± 2 °C were evaluated for tenderness by myofibrillar fragmentation index determination, shear force analysis and sensorial testing. The 24 h post-harvested intact samples were 30.6% more tender than excised samples and 41.7% more tender than control samples (p ? 0.05). The myofibrillar fragments index was 13.2% higher in intact samples than in deboned fillet (p ? 0.05) and a sensory test showed that the 24 h intact samples were of major acceptability. Our results demonstrated that tenderness was best achieved with intact breast fillet samples stored at 2 ± 2 °C for 24 h.     

钙激活酶对肉嫩度的影响

宰后肌肉嫩度的变化是在多种酶的协同作用下完成的,其中一个重要部分是钙激活酶。本文主要论述了钙激活酶的分子结构特征,影响酶活性的因素、以及对肌肉嫩化的作用机理等方面的内容。     

DEAE-Sephacel和DEAE-Sepharose-FF 对牛肉中Calpains分离纯化的比较研究

实验通过DEAE离子交换层析对牛背长肌中钙激活酶(Calpains)的分离研究,比较了DEAE-Sepharose-FF和DEAE-Sephacel对钙激活酶(Calpains)的分离效果,确定了Calpains分离纯化的条件.Sephacel不耐压,分离时间长,因此选择Sepharose-FF分离纯化Calpains.