Blue native-PAGE （BN-PAGE） resolves protein complexes in their native state. When combined with immu- noblotting, it can be used to identify the presence of high molecular weight complexes at high resolution for any...Blue native-PAGE （BN-PAGE） resolves protein complexes in their native state. When combined with immu- noblotting, it can be used to identify the presence of high molecular weight complexes at high resolution for any protein, given a suitable antibody. To identify proteins in high molecular weight complexes on a large scale and to bypass the requirement for specific antibodies, we applied a tandem mass spectrometry （MS/MS） approach to BN-PAGE-resolved chloroplasts. Fractionation of the gel into six bands allowed iden- tification and label-free quantification of 1000 chloroplast proteins with native molecular weight separation. Significantly, this approach achieves a depth of identification comparable with traditional shotgun proteo- mic analyses of chloroplasts, indicating much of the known chloroplast proteome is amenable to MS/MS identification under our fractionation scheme. By limiting the number of fractionation bands to six, we facil- itate scaled-up comparative analyses, as we demonstrate with the reticulata chloroplast mutant displaying a reticulated leaf phenotype. Our comparative proteomics approach identified a candidate interacting protein of RETICULATA as well as effects on lipid remodeling proteins, amino acid metabolic enzymes, and plastid division machinery. We additionally highlight selected proteins from each sub-compartment of the chloroplast that provide novel insight on known or hypothesized protein complexes to further illus- trate the utility of this approach. Our results demonstrate the high sensitivity and reproducibility of this technique, which is anticipated to be widely adaptable to other sub-cellular compartments.展开更多
Since ＂world literature＂ has in recent years become a topic of heated international discussion in the humanities, we need to formulate a concept of world poetics. This is for the following theoretical reasons. First...Since ＂world literature＂ has in recent years become a topic of heated international discussion in the humanities, we need to formulate a concept of world poetics. This is for the following theoretical reasons. Firstly, world poetics is a natural theoretical sublation of research achievements in the field of world literature and comparative poetics. The current dominance of Western literary theories does not do justice to the experience of the literatures and literary theories of other countries and peoples. Secondly, to date, all influential literary interpretations and theories have been produced in a Western setting. However, due to the limitations of the linguistic and cultural background of such interpretations, they cannot simultaneously cover the categories and experiences of Eastern and Western literatures and literary theories. Thirdly, Chinese scholars have a long history of research in cutting-edge Western literary theories and abundant experience in indigenous Eastern literatures and critical theory. Formulating the concept of world poetics will improve world literary theories and concepts and will remap the scenario of existing world literatures and literary theories.展开更多
文摘Blue native-PAGE （BN-PAGE） resolves protein complexes in their native state. When combined with immu- noblotting, it can be used to identify the presence of high molecular weight complexes at high resolution for any protein, given a suitable antibody. To identify proteins in high molecular weight complexes on a large scale and to bypass the requirement for specific antibodies, we applied a tandem mass spectrometry （MS/MS） approach to BN-PAGE-resolved chloroplasts. Fractionation of the gel into six bands allowed iden- tification and label-free quantification of 1000 chloroplast proteins with native molecular weight separation. Significantly, this approach achieves a depth of identification comparable with traditional shotgun proteo- mic analyses of chloroplasts, indicating much of the known chloroplast proteome is amenable to MS/MS identification under our fractionation scheme. By limiting the number of fractionation bands to six, we facil- itate scaled-up comparative analyses, as we demonstrate with the reticulata chloroplast mutant displaying a reticulated leaf phenotype. Our comparative proteomics approach identified a candidate interacting protein of RETICULATA as well as effects on lipid remodeling proteins, amino acid metabolic enzymes, and plastid division machinery. We additionally highlight selected proteins from each sub-compartment of the chloroplast that provide novel insight on known or hypothesized protein complexes to further illus- trate the utility of this approach. Our results demonstrate the high sensitivity and reproducibility of this technique, which is anticipated to be widely adaptable to other sub-cellular compartments.
基金a phased achievement of the Major Prograrr "The Study of Marxism and World Literature" funded by the National Social Science Fund(Grant No.:14ZDB082)
文摘Since ＂world literature＂ has in recent years become a topic of heated international discussion in the humanities, we need to formulate a concept of world poetics. This is for the following theoretical reasons. Firstly, world poetics is a natural theoretical sublation of research achievements in the field of world literature and comparative poetics. The current dominance of Western literary theories does not do justice to the experience of the literatures and literary theories of other countries and peoples. Secondly, to date, all influential literary interpretations and theories have been produced in a Western setting. However, due to the limitations of the linguistic and cultural background of such interpretations, they cannot simultaneously cover the categories and experiences of Eastern and Western literatures and literary theories. Thirdly, Chinese scholars have a long history of research in cutting-edge Western literary theories and abundant experience in indigenous Eastern literatures and critical theory. Formulating the concept of world poetics will improve world literary theories and concepts and will remap the scenario of existing world literatures and literary theories.
