Carbonic anhydrase accounts for catalytic reaction of CO_(2)/HCO_(3)^(–) transformation, thus resulting in neutralization and acidification of the cellular environment, thereby favoring tumor development. Hence, it i...Carbonic anhydrase accounts for catalytic reaction of CO_(2)/HCO_(3)^(–) transformation, thus resulting in neutralization and acidification of the cellular environment, thereby favoring tumor development. Hence, it is a classical protein model of greatly biocatalytic significance as well as a highly expressed biomarker with renal tumor. We herein proposed a single-molecule measurement on carbonic anhydrase using MspA nanopore, in [BMIM+] and asymmetric K^(+)/Ca^(2+) cationic coordinated environment, instead of usual symmetric KCl/NaCl electrolyte. Significantly, our empirical analysis showed that asymmetric K^(+)/Ca^(2+) cationic environment contributes to distinguishable current modulations, thus yielding better resolution for carbonic anhydrase measurement, which is independent of applied voltage and more importantly, is stable enough at varied pH conditions and for very low concentration test in urine sample. Our results provide a classical model for nanopore protein analysis, and may also permit biocatalytic measurement at single-molecule level.展开更多
Characterization of the colon cancer immunome and its autoantibody signature from differentially-reactive antigens （DIRAGs） could provide insights into aberrant cellular mechanisms or enriched networks associated wi...Characterization of the colon cancer immunome and its autoantibody signature from differentially-reactive antigens （DIRAGs） could provide insights into aberrant cellular mechanisms or enriched networks associated with diseases. The purpose of this study was to characterize the antibody profile of plasma samples from 32 colorectal cancer （CRC） patients and 32 controls using proteins isolated from 15,417 human cDNA expression clones on microarrays. 671 unique DIRAGs were identified and 632 were more highly reactive in CRC samples. Bioinformatics analyses reveal that compared to control samples, the immunoproteomic IgG profiling of CRC samples is mainly associated with cell death, survival, and proliferation pathways, especially proteins involved in EIF2and mTOR signaling. Ribosomal proteins （e.g., RPL7, RPL22, and RPL27A） and CRC-related genes such as APC, AXIN1, E2F4, MSH2, PMS2, and TP53 were highly enriched. In addition, dif- ferential pathways were observed between the CRC and control samples. Furthermore, 103 DIR- AGs were reported in the SEREX antigen database, demonstrating our ability to identify known and new reactive antigens. We also found an overlap of 7 antigens with 48 ＂CRC genes.＂ These data indicate that immunomies profiling on protein mieroarrays is able to reveal the complexity of immune responses in cancerous diseases and faithfully reflects the underlying pathology.展开更多
基金National Key Research and Development Program of China(2022YFB3205600)Chongqing Talents:Exceptional Young Talents Project(cstc2021ycjh-bgzxm0016)+1 种基金Beibei Technology Talents and Independent Innovation Project(No.2022-34)the Natural Science Foundation of Chongqing,China(cstc2021jcyj-jqx0030).
文摘Carbonic anhydrase accounts for catalytic reaction of CO_(2)/HCO_(3)^(–) transformation, thus resulting in neutralization and acidification of the cellular environment, thereby favoring tumor development. Hence, it is a classical protein model of greatly biocatalytic significance as well as a highly expressed biomarker with renal tumor. We herein proposed a single-molecule measurement on carbonic anhydrase using MspA nanopore, in [BMIM+] and asymmetric K^(+)/Ca^(2+) cationic coordinated environment, instead of usual symmetric KCl/NaCl electrolyte. Significantly, our empirical analysis showed that asymmetric K^(+)/Ca^(2+) cationic environment contributes to distinguishable current modulations, thus yielding better resolution for carbonic anhydrase measurement, which is independent of applied voltage and more importantly, is stable enough at varied pH conditions and for very low concentration test in urine sample. Our results provide a classical model for nanopore protein analysis, and may also permit biocatalytic measurement at single-molecule level.
基金supported by the Life Science Krems Fund (Project No. 30)Jubilumsfonds of the Austrian National Bank (Project No. 15192)Vienna Science and Technology Fund (Project No LS11-026) of Austria
文摘Characterization of the colon cancer immunome and its autoantibody signature from differentially-reactive antigens （DIRAGs） could provide insights into aberrant cellular mechanisms or enriched networks associated with diseases. The purpose of this study was to characterize the antibody profile of plasma samples from 32 colorectal cancer （CRC） patients and 32 controls using proteins isolated from 15,417 human cDNA expression clones on microarrays. 671 unique DIRAGs were identified and 632 were more highly reactive in CRC samples. Bioinformatics analyses reveal that compared to control samples, the immunoproteomic IgG profiling of CRC samples is mainly associated with cell death, survival, and proliferation pathways, especially proteins involved in EIF2and mTOR signaling. Ribosomal proteins （e.g., RPL7, RPL22, and RPL27A） and CRC-related genes such as APC, AXIN1, E2F4, MSH2, PMS2, and TP53 were highly enriched. In addition, dif- ferential pathways were observed between the CRC and control samples. Furthermore, 103 DIR- AGs were reported in the SEREX antigen database, demonstrating our ability to identify known and new reactive antigens. We also found an overlap of 7 antigens with 48 ＂CRC genes.＂ These data indicate that immunomies profiling on protein mieroarrays is able to reveal the complexity of immune responses in cancerous diseases and faithfully reflects the underlying pathology.
