TY - JOUR
T1 - Characterization of In Vivo Protein Complexes via Chemical Cross-Linking and Mass Spectrometry
AU - Wang, Yuefan
AU - Hu, Yingwei
AU - Höti, Naseruddin
AU - Huang, Lan
AU - Zhang, Hui
N1 - Publisher Copyright:
© 2021 American Chemical Society
PY - 2022/1/25
Y1 - 2022/1/25
N2 - Cells perform various functions by proteins via protein complexes. Characterization of protein complexes is critical to understanding their biological and clinical significance and has been one of the major efforts of functional proteomics. To date, most protein complexes are characterized by the in vitro system from protein extracts after the cells or tissues are lysed, and it has been challenging to determine which of these protein complexes are formed in intact cells. Herein, we report an approach to preserve protein complexes using in vivo cross-linking, followed by size exclusion chromatography and data-independent acquisition mass spectrometry. This approach enables the characterization of in vivo protein complexes from cells or tissues, which allows the determination of protein complexes in clinical research. More importantly, the described approach can identify protein complexes that are not detected by the in vitro system, which provide unique protein function information.
AB - Cells perform various functions by proteins via protein complexes. Characterization of protein complexes is critical to understanding their biological and clinical significance and has been one of the major efforts of functional proteomics. To date, most protein complexes are characterized by the in vitro system from protein extracts after the cells or tissues are lysed, and it has been challenging to determine which of these protein complexes are formed in intact cells. Herein, we report an approach to preserve protein complexes using in vivo cross-linking, followed by size exclusion chromatography and data-independent acquisition mass spectrometry. This approach enables the characterization of in vivo protein complexes from cells or tissues, which allows the determination of protein complexes in clinical research. More importantly, the described approach can identify protein complexes that are not detected by the in vitro system, which provide unique protein function information.
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U2 - 10.1021/acs.analchem.1c02410
DO - 10.1021/acs.analchem.1c02410
M3 - Article
C2 - 34962381
AN - SCOPUS:85122446469
SN - 0003-2700
VL - 94
SP - 1537
EP - 1542
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 3
ER -