采用固相pH梯度-SDS PAGE双向电泳对红莲型细胞质雄性不育水稻的不育系(YTA)和保持系(YTB)单核期花粉总蛋白质进行了分离,通过银染显色,获得了分辨率和重复性较好的双向电泳图谱。Image Master 2DV5.0软件可识别约1800个蛋白质点,其中...采用固相pH梯度-SDS PAGE双向电泳对红莲型细胞质雄性不育水稻的不育系(YTA)和保持系(YTB)单核期花粉总蛋白质进行了分离,通过银染显色,获得了分辨率和重复性较好的双向电泳图谱。Image Master 2DV5.0软件可识别约1800个蛋白质点,其中差异表达的蛋白质点数为85。将其中16个差异点采用基质辅助激光解析电离飞行时间质谱(matrix assisted laser desorption/ionizaton time of flight mass spectrometry,MALDI-TOF-MS)进行了肽质指纹图分析,通过采用Mascot软件对MSDB数据库查询,其中9个蛋白质点得到了鉴定。YTA相对于YTB有部分参与碳代谢和淀粉合成的酶缺失或表达量降低,这些蛋白质分别是ADP-葡萄糖磷酸转移酶(AGPase),UDP-葡萄糖醛酸脱羧酶,乙酰辅酶A合成酶和二氢硫辛酸脱氢酶等。其中AGPase是参与淀粉合成的蛋白,与花粉发育密切相关。乙酰辅酶A合成酶和二氢硫辛酸脱氢酶是细胞内合成乙酰辅酶A的重要酶,而乙酰辅酶A是进入TCA循环的重要底物,乙酰辅酶A的缺乏可以导致TCA循环不能顺利进行,从而不能提供小孢子发育所需要的大量能量。YTA相对于YTB部分参与碳水化合物代谢的重要酶缺失或表达量降低,有可能导致因线粒体提供的能量不足,淀粉合成受阻,因而花粉不能正常发育。展开更多
AIM： To detect the new serum biomarkers for colorectal cancer （CRC） by serum protein profiling with surfaceenhanced laser desorption ionisation - time of flight mass spectrometry （SELDI-TOF MS）. METHODS： Two ind...AIM： To detect the new serum biomarkers for colorectal cancer （CRC） by serum protein profiling with surfaceenhanced laser desorption ionisation - time of flight mass spectrometry （SELDI-TOF MS）. METHODS： Two independent serum sample sets were analysed separately with the ProteinChip technology （set A： 40 CRC ＋ 49 healthy controls; set B： 37 CRC ＋ 31 healthy controls）, using chips with a weak cation exchange moiety and buffer pH 5. Discriminative power of differentially expressed proteins was assessed with a classification tree algorithm. Sensitivities and specificities of the generated classification trees were obtained by blindly applying data from set A to the generated trees from set B and vice versa. CRC serum protein profiles were also compared with those from breast, ovarian, prostate, and non-small cell lung cancer. RESULTS： Mass-to-charge ratios （m/z） 3.1×10^3, 3.3× 10^3, 4.5×10^3, 6.6×10^3 and 28×10^3 were used as classitiers in the best-performing classification trees. Tree sensitivities and specificities were between 65% and 90%.Host of these discriminative m/z values were also different in the other tumour types investigated. M/z 3.3× 10^3, main classifier in most trees, was a doubly charged form of the 6.6× 10^3-Da protein. The latter was identified as apolipoprotein C-I. M/z 3.1×10^3 was identified as an N-terminal fragment of albumin, and m/z 28× 10^3 as apolipoprotein A-I. CONCLUSION： SELDI-TOF MS followed by classification tree pattern analysis is a suitable technique for finding new serum markers for CRC. Biomarkers can be identified and reproducibly detected in independent sample sets with high sensitivities and specificities. Although not specific for CRC, these biomarkers have a potential role in disease and treatment monitoring.展开更多
文摘采用固相pH梯度-SDS PAGE双向电泳对红莲型细胞质雄性不育水稻的不育系(YTA)和保持系(YTB)单核期花粉总蛋白质进行了分离,通过银染显色,获得了分辨率和重复性较好的双向电泳图谱。Image Master 2DV5.0软件可识别约1800个蛋白质点,其中差异表达的蛋白质点数为85。将其中16个差异点采用基质辅助激光解析电离飞行时间质谱(matrix assisted laser desorption/ionizaton time of flight mass spectrometry,MALDI-TOF-MS)进行了肽质指纹图分析,通过采用Mascot软件对MSDB数据库查询,其中9个蛋白质点得到了鉴定。YTA相对于YTB有部分参与碳代谢和淀粉合成的酶缺失或表达量降低,这些蛋白质分别是ADP-葡萄糖磷酸转移酶(AGPase),UDP-葡萄糖醛酸脱羧酶,乙酰辅酶A合成酶和二氢硫辛酸脱氢酶等。其中AGPase是参与淀粉合成的蛋白,与花粉发育密切相关。乙酰辅酶A合成酶和二氢硫辛酸脱氢酶是细胞内合成乙酰辅酶A的重要酶,而乙酰辅酶A是进入TCA循环的重要底物,乙酰辅酶A的缺乏可以导致TCA循环不能顺利进行,从而不能提供小孢子发育所需要的大量能量。YTA相对于YTB部分参与碳水化合物代谢的重要酶缺失或表达量降低,有可能导致因线粒体提供的能量不足,淀粉合成受阻,因而花粉不能正常发育。
文摘AIM： To detect the new serum biomarkers for colorectal cancer （CRC） by serum protein profiling with surfaceenhanced laser desorption ionisation - time of flight mass spectrometry （SELDI-TOF MS）. METHODS： Two independent serum sample sets were analysed separately with the ProteinChip technology （set A： 40 CRC ＋ 49 healthy controls; set B： 37 CRC ＋ 31 healthy controls）, using chips with a weak cation exchange moiety and buffer pH 5. Discriminative power of differentially expressed proteins was assessed with a classification tree algorithm. Sensitivities and specificities of the generated classification trees were obtained by blindly applying data from set A to the generated trees from set B and vice versa. CRC serum protein profiles were also compared with those from breast, ovarian, prostate, and non-small cell lung cancer. RESULTS： Mass-to-charge ratios （m/z） 3.1×10^3, 3.3× 10^3, 4.5×10^3, 6.6×10^3 and 28×10^3 were used as classitiers in the best-performing classification trees. Tree sensitivities and specificities were between 65% and 90%.Host of these discriminative m/z values were also different in the other tumour types investigated. M/z 3.3× 10^3, main classifier in most trees, was a doubly charged form of the 6.6× 10^3-Da protein. The latter was identified as apolipoprotein C-I. M/z 3.1×10^3 was identified as an N-terminal fragment of albumin, and m/z 28× 10^3 as apolipoprotein A-I. CONCLUSION： SELDI-TOF MS followed by classification tree pattern analysis is a suitable technique for finding new serum markers for CRC. Biomarkers can be identified and reproducibly detected in independent sample sets with high sensitivities and specificities. Although not specific for CRC, these biomarkers have a potential role in disease and treatment monitoring.
