Identification of the core motif of the BRCA2 C-terminal RAD51-binding domain by comparing canine and human BRCA2

Mammary tumors are the most common tumors in women and non-spayed female dogs. One of the reasons for mammary tumors is mutations of the tumor suppressor gene, BRCA2. BRCA2 participates in homologous recombination repair by interacting with the RAD51 recombinase. BRCA2 has two RAD51-binding domains, consisting of BRC repeats and the C-terminal RAD51-binding domain, respectively. Although several studies have addressed the function of the C-terminal RAD51-binding domain of human BRCA2, the amino acid sequences required for the RAD51-interaction activity remain unclear. In this study, the C-terminal RAD51-binding domains of canine and human BRCA2 were compared; the canine domain displayed a weaker interaction with RAD51. This difference was attributed to the C-terminal portion of the domain via a comparison between canine and human domains. Furthermore, peptides shorter than those previously reported displayed RAD51-interacting activity, and a core motif of this domain consisting of 25 amino acids was identified. Since a mutation (S3323N) was reported in the core motif of this domain, the effect of this mutation was evaluated. The mutant exhibited similar RAD51-binding activity as that of the wild-type protein, suggesting that the mutation was functionally neutral. These data suggested that the C-terminal portion of the BRCA2 C-terminal RAD51-binding domain influenced its RAD51-interaction activity, and a minimum core motif of 25 amino acids was identified in this domain. These data may help clarify BRCA2 function, as well as the tumorigenic effects of BRCA2 mutation.     

自溶法回收鱼类加工废弃物中蛋白质成分(英文)

鱼头和鱼内脏为鱼类在加工过程中废弃的主要组织,其中含有丰富的蛋白质和脂肪等有益成分,有必要进行回收利用。双鳍舵鲣鱼的头部和内脏约占其鱼体重量的30％,富含蛋白质(14.4％-22.6％)。为了将这些废弃物中的蛋白质进行回收,分别采用双鳍舵鲣鱼内脏本身含有的内源性蛋白酶(自溶)和四种外源性的商业蛋白酶Papain W-40(E1),Pancidase NP-2(E2),Sumichiimu FP(E3)和Protease M(E4)对废弃物中的蛋白质进行水解(50℃,1～4h),考察自溶和不同酶解方法对蛋白质的回收率以及对回收液氨基酸组成的影响。结果显示,自溶法对废弃物中蛋白质的回收效果最好。当水解温度为50℃,水解时间为2h时,自溶法的蛋白质回收率最高,达到76％;其次为E2酶解法,回收率达到70％;E3酶解法效果最差,回收率为63％。进一步研究表明,自溶回收液(AE)的氨基酸总量也最高,达到11756 mg/100g,而采用E1、E2、E3和E4水解方法得到的酶解回收液(EE1、EE2、EE3和EE4)中氨基酸含量分别仅为10294、10642、8750、10121 mg/100g。感观评定结果表明,自溶回...     

Lipid composition and deposition of cultured yellowtail Seriola quinqueradiata muscle at different anatomical locations in relation to meat texture

ABSTRACT:   The present study was undertaken to assess the lipid composition and deposition in muscle at three anatomical locations in cultured yellowtail and to investigate the effect of lipid composition and deposition on meat texture. Lipid deposition in muscle was studied by histochemical staining of lipid with Sudan dye. Lipid class composition analysis showed that neutral lipids were the main constituents of lipid in cultured yellowtail and accounted primarily for the variation in muscle lipid content with the anatomical location of meat, as well as with season, whereas the polar lipid content remained almost constant. Furthermore, muscle neutral lipid content was correlated negatively with meat breaking strength; however, no correlation was observed between muscle polar lipid content and meat breaking strength. The histochemical study revealed that, in yellowtail muscle, lipid is preferentially deposited in the myosepta and, with increases in muscle lipid content, additional fat is deposited along sparsely distributed thin connective tissue. It was also observed that the greater the lipid deposition in collageneous connective tissue, the lower the meat breaking strength; presumably, higher lipid deposition in the connective tissue resulted in weakening of the muscle structure.     

自溶法回收鱼类加工废弃物中蛋白质成分

鱼头和鱼内脏为鱼类在加工过程中废弃的主要组织,其中含有丰富的蛋白质和脂肪等有益成分,有必要进行回收利用。双鳍舵鲣鱼的头部和内脏约占其鱼体重量的30％,富含蛋白质(14.4％-22.6％)。为了将这些废弃物中的蛋白质进行回收,分别采用双鳍舵鲣鱼内脏本身含有的内源性蛋白酶(自溶)和四种外源性的商业蛋白酶Papain W-40(E1),Pancidase NP-2(E2),Sumichiimu FP(E3)和Protease M(E4)对废弃物中的蛋白质进行水解(50℃,1～4h),考察自溶和不同酶解方法对蛋白质的回收率以及对回收液氨基酸组成的影响。结果显示,自溶法对废弃物中蛋白质的回收效果最好。当水解温度为50℃,水解时间为2h时,自溶法的蛋白质回收率最高,达到76％;其次为E2酶解法,回收率达到70％;E3酶解法效果最差,回收率为63％。进一步研究表明,自溶回收液(AE)的氨基酸总量也最高,达到11756 mg/100g,而采用E1、E2、E3和E4水解方法得到的酶解回收液(EE1、EE2、EE3和EE4)中氨基酸含量分别仅为10294、10642、8750、10121 mg/100g。感观评定结果表明,自溶回...     

Gel forming ability of fish meat oxidized during washing

ABSTRACT: In order to examine the effect of meat oxidation on the gel forming ability before grinding the meat with salt, fish meat was washed with CuCl₂ solution, and the gel strength as well as total sulfhydryl (SH) groups and sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE) patterns were analysed. Washing with CuCl₂ solution resulted in a decrease in the total SH content of fish meat and the formation of myosin heavy chain (MHC) dimer through disulfide bonding. The plot of logarithmic gel strength versus protein concentration after heating the washed meat at 80°C in the presence of 3% NaCl to form a gel illustrated that the gel forming ability of meats washed with CuCl₂ solution was weaker than the control meat. The gel of meat washed with CuCl₂ showed the polymerization of MHC and MHC dimers through disulfide bonding much more than the control meat gel, although a small decrease in the SH group content after heating. Further washing with ethylene diamine tetra-acetic acid (EDTA) solution to remove CuCl₂ from the CuCl₂-washed meat also resulted in similar behavior for MHC polymerization and SH content as the CuCl₂-washed meat, and the gel was still weaker than the control gel. It was found that the oxidation of SH groups during washing with CuCl₂ solution accompanied by MHC dimer formation in the meat results in the weakening of its gel forming ability.