Catalytically Inactive SHP1-C453S Mutant Gain of “Robust LLPS” Function
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Abstract
SHP1 is a non-receptor protein tyrosine phosphatase extensively expressed in hematopoietic cells, exerting a pivotal role as an immunosuppressive factor. Our previous studies have suggested that SHP1 can undergo liquid-liquid phase separation (LLPS). In this study, the SHP1-C455S mutant, commonly utilized in biochemical assays due to its lack of catalytic phosphatase activity, unexpectedly exhibited a remarkably robust ability for LLPS. Since the C453S mutation has been previously shown to potentially induce a conformational transition of SHP1 from a closed to an open state, we hypothesize that the enhanced LLPS capability of SHP1 may be facilitated by this conformational alteration. The SHP1-C453S mutant exhibited robust LLPS activity, while completely abrogating its phosphatase activity.
This allows for effective investigation of the catalytic activity and LLPS capability of SHP1.
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Matthews RJ, Bowne DB, Flores E, Thomas ML. Characterization of hematopoietic intracellular protein tyrosine phosphatases: description of a phosphatase containing an SH2 domain and another enriched in proline-, glutamic acid-, serine-, and threonine-rich sequences. Mol Cell Biol. 1992;12(5):2396-2405. Available from: https://doi.org/10.1128/mcb.12.5.2396-2405.1992
Yi TL, Cleveland JL, Ihle JN. Protein tyrosine phosphatase containing SH2 domains: characterization, preferential expression in hematopoietic cells, and localization to human chromosome 12p12-p13. Mol Cell Biol. 1992;12(2):836-846. Available from: https://doi.org/10.1128/mcb.12.2.836-846.1992
Tsui FWL, Martin A, Wang J, Tsui HW. Investigations into the regulation and function of the SH2 domain-containing protein-tyrosine phosphatase, SHP-1. Immunol Res. 2006;35(1-2):127-136. Available from: https://doi.org/10.1385/ir:35:1:127
Garg M, Wahid M, Khan F. Regulation of peripheral and central immunity: understanding the role of Src homology 2 domain-containing tyrosine phosphatases, SHP-1 & SHP-2. Immunobiology. 2020;225(1):151847. Available from: https://doi.org/10.1016/j.imbio.2019.09.006
Abram CL, Lowell CA. Shp1 function in myeloid cells. J Leukoc Biol. 2017;102(3):657-675. Available from: https://doi.org/10.1189/jlb.2mr0317-105r
Mehta S, Zhang J. Liquid-liquid phase separation drives cellular function and dysfunction in cancer. Nat Rev Cancer. 2022;22(4):239-252. Available from: https://doi.org/10.1038/s41568-022-00444-7
Hirose T, Ninomiya K, Nakagawa S, Yamazaki T. A guide to membraneless organelles and their various roles in gene regulation. Nat Rev Mol Cell Biol. 2023;24(4):288-304. Available from: https://doi.org/10.1038/s41580-022-00558-8
Alberti S, Gladfelter A, Mittag T. Considerations and challenges in studying liquid-liquid phase separation and biomolecular condensates. Cell. 2019;176(3):419-434. Available from: https://doi.org/10.1016/j.cell.2018.12.035
Banani SF, Lee HO, Hyman AA, Rosen MK. Biomolecular condensates: organizers of cellular biochemistry. Nat Rev Mol Cell Biol. 2017;18(5):285-298. Available from: https://doi.org/10.1038/nrm.2017.7
Zhang Q, Kong W, Zhu T, Zhu G, Zhu J, Kong X, et al. Phase separation ability and phosphatase activity of the SHP1-R360E mutant. Biochem Biophys Res Commun. 2022;600:150-155. Available from: https://doi.org/10.1016/j.bbrc.2022.02.070
Lin S, Raval S, Zhang Z, Deverill M, Siminovitch KA, Branch DR, et al. The protein-tyrosine phosphatase SHP-1 regulates the phosphorylation of alpha-actinin. J Biol Chem. 2004;279(24):25755-25764. Available from: https://doi.org/10.1074/jbc.m314175200
Dustin L, Plas D, Wong J, Hu YT, Soto C, Chan AC, et al. Expression of dominant-negative src-homology domain 2-containing protein tyrosine phosphatase-1 results in increased Syk tyrosine kinase activity and B cell activation. J Immunol (Baltimore, Md: 1950). 1999;162(5):2717-2724. Available from: https://pubmed.ncbi.nlm.nih.gov/10072516/
Sha F, Kurosawa K, Glasser E, Ketavarapu G, Albazzaz S, Koide A, et al. Monobody inhibitor selective to the phosphatase domain of SHP2 and its use as a probe for quantifying SHP2 allosteric regulation. J Mol Biol. 2023;435(8):168010. Available from: https://doi.org/10.1016/j.jmb.2023.168010
Zhu G, Xie J, Kong W, Xie J, Li Y, Du L, et al. Phase separation of disease-associated SHP2 mutants underlies MAPK hyperactivation. Cell. 2020;183(2):490-502.e18. Available from: https://doi.org/10.1016/j.cell.2020.09.002
Yang J, Liu L, He D, Song X, Liang X, Zhao ZJ, et al. Crystal structure of human protein-tyrosine phosphatase SHP-1. J Biol Chem. 2003;278(8):6516-6520. Available from: https://doi.org/10.1074/jbc.m210430200
Wang W, Liu L, Song X, Mo Y, Komma C, Bellamy HD, et al. Crystal structure of human protein tyrosine phosphatase SHP-1 in the open conformation. J Cell Biochem. 2011;112(8):2062-2071. Available from: https://doi.org/10.1002/jcb.23125
Tamura A, Ito G, Matsuda H, Nibe-Shirakihara Y, Hiraoka Y, Kitagawa S, et al. Zranb1-mutant mice display abnormal colonic mucus production and exacerbation of DSS-induced colitis. Biochem Biophys Res Commun. 2022;628:147-154. Available from: https://doi.org/10.1016/j.bbrc.2022.08.046