Postmortem titin proteolysis is influenced by sarcomere length in bovine muscle

The calpain protease system, in particular, μ-calpain is involved in the disassembly of specific myofibrillar proteins, resulting in tenderization of meat postmortem. Given the size, complexity, and integral nature of titin to the structure of the sarcomere, it is plausible that the length of a sarcomere may alter the susceptibility of various domains of titin to cleavage by the calpains. Therefore, we hypothesized titin degradation differs in a sarcomere-length-dependent manner in beef. After slaughter, beef carcasses were split and sides were either suspended by the Achilles tendon (normal suspension, NS) or by the aitchbone (hip suspension, HS). Immediately after suspension, samples were dissected from the LM, psoas major (PM), and semitendinosus (STN) muscles to serve as 0-d controls. After 24 h, 4 steaks were removed from each muscle and randomly assigned to 1-, 4-, 7-, or 10-d aging treatments. After the assigned aging period, myofibrils were purified for determination of sarcomere length. Warner-Bratzler shear force analysis was also performed to evaluate differences in tenderness. Muscle proteins were solubilized and subjected to SDS-VAGE (vertical agarose gel electrophoresis) to evaluate titin degradation. Sarcomere lengths differed (P < 0.0001) between contralateral muscles of NS and HS carcasses. Quantification of SDS-VAGE gels revealed less (P < 0.05) intact titin in the PM muscle of NS carcasses at each aging period compared with the PM of HS carcasses. No significant differences (P > 0.05) were detected in the disappearance of intact titin among suspension methods in the LM or STN. These data demonstrate that suspension method alters proteolysis of titin and suggest an increase in sarcomere length may contribute to the susceptibility of titin to postmortem proteolysis in beef.     

与土壤盐碱性有关的水稻生产问题

最近阿肯色大学承担了“与土壤盐碱有关的水稻和大豆生产上存在的问题”的研究课题 ,目的是为深入研究和建立最佳管理模型奠定基础。过去在水稻生产问题的性质和定义方面 ,许多概念含糊不清。因此 ,“生产问题”的准确描述对于科学家、咨询机构及种植者了解、研究和解决这些问题非常必要。1　问题的确定在传统水稻———大豆轮作过程中遇到的问题起初都被定义为“与土壤有关的问题”。由于连续灌溉 ,许多所谓的“土壤问题”实质上是经常使用劣质水灌溉的一种直接结果。大多数问题表现在有机质含量低的粉砂或砂壤土上。这些土壤缓冲性很低 ,…     

Dimerization of the extracellular domain of the human growth hormone receptor by a single hormone molecule

Human growth hormone (hGH) forms a 1:2 complex with the extracellular domain of its receptor-binding protein (hGHbp) as studied by crystallization, size exclusion chromatography, calorimetry, and a previously undescribed fluorescence quenching assay. These and other experiments with protein engineered variants of hGH have led to the identification of the binding determinants for two distinct but adjacent sites on hGH for the hGHbp, and the data indicated that there are two overlapping binding sites on the hGHbp for hGH. Furthermore, the binding of hGH to the hGHbp occurred sequentially; a first hGHbp molecule bound to site 1 on hGH and then a second hGHbp bound to site 2. Hormone-induced receptor dimerization is proposed to be relevant to the signal transduction mechanism for the hGH receptor and other related cytokine receptors.     

Glycosylation of nucleocytoplasmic proteins: signal transduction and O-GlcNAc

The dynamic glycosylation of serine or threonine residues on nuclear and cytosolic proteins by O-linked beta-N-acetylglucosamine (O-GlcNAc) is abundant in all multicellular eukaryotes. On several proteins, O-GlcNAc and O-phosphate alternatively occupy the same or adjacent sites, leading to the hypothesis that one function of this saccharide is to transiently block phosphorylation. The diversity of proteins modified by O-GlcNAc implies its importance in many basic cellular and disease processes. Here we systematically examine the current data implicating O-GlcNAc as a regulatory modification important to signal transduction cascades.     

Blood oxygen transport and hemoglobin function in three tropical fish species from northern Australian freshwater billabongs

The oxygen binding properties of whole blood and hemoglobin were studied in three phylogenetically distant tropical teleosts inhabiting freshwater billabongs that exhibit marked seasonal oxygen stratification. The water breathing saratoga (Scleropages jardinii, Family Osteoglossidae), derived from an ancient divergence of the stem line leading to all other extant teleosts, has a low blood oxygen carrying capacity, Bohr and Root effect, and a high blood oxygen affinity. Erythrocytes contain a single major hemoglobin component, and equimolar concentrations of ATP and GTP. The tarpon (Megalops cyprinoides, Family Megalopidae), a facultative air-breather, has the highest blood oxygen carrying capacity and Bohr effect, and a low blood oxygen affinity and Root effect. Erythrocytes contain a single major hemoglobin and ATP as a putative regulator of hemoglobin oxygen affinity. Barramundi (Lates calcarifer, Family Centropomidae), is an obligate water-breather with intermediate blood oxygen binding properties, and the smallest Root effect. Erythrocytes contain at least 7 hemoglobins, and equimolar concentrations of ATP and GTP. Functional properties of these three blood oxygen transport systems are considered in terms of the respiratory environment and demand for oxygen. Our interpretation supports the hypothesis that the process of speciation can lead to divergence in physiological mechanisms, irrespective of past or present selection pressures.     

Fish biodiversity sampling in stream ecosystems: a process for evaluating the appropriate types and amount of gear

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Inclusion of camelina meal as a protein source in diets for farmed Atlantic cod Gadus morhua

Camelina meal Camelina sativa (CM) is a potential protein source in aquaculture feeds, because of its crude protein level (39%) and essential amino acids. Two feeding experiments were conducted with Atlantic cod Gadus morhua. Cod in Experiment I (19.4 g fish^?1) were fed diets with 0%, 12% or 24% CM for 9.5 weeks at 10°C; and cod in Experiment II (14.4 g fish^?1) were fed diets with 0%, 15%, 30% or 40% CM for 13 weeks at 10°C. Growth, lipid and amino acid tissue composition were compared amongst cod fed varying levels of CM. In Experiment I, cod could tolerate the highest level of CM inclusion (24%) without affecting growth compared to cod fed the control diet. In Experiment II, growth performance was significantly affected at 30% CM inclusion compared to the control treatment, and cod fed 15% CM displayed some signs of depressed growth (reduced feed intake and weight gain). Both treatment and duration were interacting factors (P = 0.015) that determined growth performance when comparing both experiments. Muscle tissue composition was relatively unaltered with less than 30% CM inclusion; however, multivariate statistics revealed significant differences in muscle tissue fatty acid composition between cod fed 40% CM and the control diet. The tissue amino acid profile was generally unaltered because the dietary amino acid profile was consistent after CM inclusion. A few antinutritive compounds in CM may have affected palatability in diets with greater than 30% CM inclusion, which may have resulted in reduced growth performance.     

Galactose toxicity in the chick: hyperosmolality

Galactose-fed chicks have been found to develop severe hyperosmolar dehydration. Although biochemical abnormalities have been observed in the brain of the galactose-toxic chick, the observed physiologic alteration of serum osmolality could be the major factor responsible for the galactose toxicity syndrome in the chick.     

Engineering enzyme specificity by "substrate-assisted catalysis"

A novel approach to engineering enzyme specificity is presented in which a catalytic group from an enzyme is first removed by site-directed mutagenesis causing inactivation. Activity is then partially restored by substrates containing the missing catalytic functional group. Replacement of the catalytic His with Ala in the Bacillus amyloliquefaciens subtilisin gene (the mutant is designated His64Ala) by site-directed mutagenesis reduces the catalytic efficiency (kcat/Km) by a factor of a million when assayed with N-succinyl-L-Phe-L-Ala-L-Ala-L-Phe-p-nitroanilide (sFAAF-pNA). Model building studies showed that a His side chain at the P2 position of a substrate bound at the active site of subtilisin could be virtually superimposed on the catalytic His side chain of this serine protease. Accordingly, the His64Ala mutant hydrolyzes a His P2 substrate (sFAHF-pNA) up to 400 times faster than a homologous Ala P2 or Gln P2 substrate (sFAAF-pNA or sFAQF-pNA) at pH 8.0. In contrast, the wild-type enzyme hydrolyzes these three substrates with similar catalytic efficiencies. Additional data from substrate-dependent pH profiles and hydrolysis of large polypeptides indicate that the His64Ala mutant enzyme can recover partially the function of the lost catalytic histidine from a His P2 side chain on the substrate. Such "substrate-assisted catalysis" provides a new basis for engineering enzymes with very narrow and potentially useful substrate specificities. These studies also suggest a possible functional intermediate in the evolution of the catalytic triad of serine proteases.