Techniques for detecting protein-protein interactions in living cells: principles, limitations, and recent progress

Cui, Yaning; Zhang, Xi; Yu, Meng; Zhu, Yingfang; Xing, Jingjing; Lin, Jinxing

doi:10.1007/s11427-018-9500-7

Cited by 57 publications

(51 citation statements)

References 98 publications

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“…However, knowledge of the function and regulation of coupled trafficking in plants remains limited compared with recent findings highlighting the mechanisms for balance and tight coupling of exo–endocytosis in animal systems. Advances in the patch clamp technique, and especially single‐molecule techniques, including total internal reflection fluorescence microscopy and super‐resolution microscopy, offer new opportunities in this field (Li et al ., ; Cui et al ., ). Although single‐molecule imaging has provided insights into factors governing endocytosis and dynamics of functional PM‐resident proteins in plants (Li et al ., ), modified imaging and analysis methods for exocytic trafficking or exo–endocytosis coupling will need to be developed.…”

Section: Concluding Remarks and Future Challengesmentioning

confidence: 97%

At the intersection of exocytosis and endocytosis in plants

2019

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Vesicle exocytosis and endocytosis control the activities and turnover of plasma membrane proteins required for signaling triggering or attenuating at the cell surface. In recent years, the diverse exocytic and endocytic trafficking pathways have been uncovered in plants. The balance between conventional and unconventional protein secretion provides an efficient strategy to respond to stress conditions. Similarly, clathrin-dependent and -independent endocytosis cooperatively regulate the dynamics of membrane proteins in response to environmental cues. In fact, many aspects of plant growth and development, such as tip growth, immune response, and protein polarity establishment, involve the tight deployment of exo-endocytic trafficking. However, our understanding of their intersection is limited. Here, we discuss recent advances in the molecular factors coupling plant exo-endocytic trafficking and the biological significance of balance between exocytosis and endocytosis in plants.

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Section: Concluding Remarks and Future Challengesmentioning

confidence: 97%

At the intersection of exocytosis and endocytosis in plants

2019

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“…, 2013; Wang et al , 2018b; Cui et al . , 2019), will enable quantification of the dynamics and assembly of the SYR1 receptor and the identification of its possible interacting partners on the membrane. This could facilitate the elucidation of the SYS‐mediated signal transduction pathways and shed new light on the mechanisms underlying SYS‐mediated innate immunity in plants.…”

Section: Concluding Remarks and Future Prospectsmentioning

confidence: 99%

Systemin‐mediated long‐distance systemic defense responses

Zhang

Lin

2020

New Phytologist

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Systemin, a peptide plant hormone of 18 amino acids, coordinates local and systemic immune responses. The activation of the canonical systemin-mediated systemic signaling pathway involves systemin release from its precursor prosystemin, systemin binding to its membrane receptor SYSTEMIN RECEPTOR1 (SYR1), and the transport of long-distance signaling molecules, including jasmonic acid, the prosystemin mRNA, volatile organic compounds and possibly systemin itself. Here, we review emerging evidence that the disordered structure and unconventional processing and secretion of systemin contribute to the regulation of systeminmediated signaling during plant defense. We highlight recent advances in systemin research, which elucidated how cells integrate multiple long-distance signals into the systemic defense response. In addition, we discuss the perception of systemin by SYR1 and its mediation of downstream defense responses.

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“…In FRET-FLIM we measure the donor's excited state decay in the presence of an acceptor [77], i.e., estimate the change in the donor's lifetime induced by the acceptor. The main advantage of the FRET-FLIM method, compared to traditional biochemical techniques for monitoring of PPIs, such as co-immunoprecipitation (co-IP), is that it supports in vivo studies, is highly sensitive and specific, and can provide information on the location of the interaction [78]. In this study, we used EGFP (the donor molecule) and mCherry (the acceptor molecule) for labeling NME proteins.…”

Section: Choosing the Optimal Fret Reporter Orientation For Estimatinmentioning

confidence: 99%

The Subcellular Localization and Oligomerization Preferences of NME1/NME2 upon Radiation-Induced DNA Damage

Radić

Šoštar

Weber

et al. 2020

IJMS

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Nucleoside diphosphate kinases (NDPK/NME/Nm23) are enzymes composed of subunits NME1/NDPK A and NME2/NDPK B, responsible for the maintenance of the cellular (d)NTP pool and involved in other cellular processes, such as metastasis suppression and DNA damage repair. Although eukaryotic NDPKs are active only as hexamers, it is unclear whether other NME functions require the hexameric form, and how the isoenzyme composition varies in different cellular compartments. To examine the effect of DNA damage on intracellular localization of NME1 and NME2 and the composition of NME oligomers in the nucleus and the cytoplasm, we used live-cell imaging and the FRET/FLIM technique. We showed that exogenous NME1 and NME2 proteins co-localize in the cytoplasm of non-irradiated cells, and move simultaneously to the nucleus after gamma irradiation. The FRET/FLIM experiments imply that, after DNA damage, there is a slight shift in the homomer/heteromer balance between the nucleus and the cytoplasm. Collectively, our results indicate that, after irradiation, NME1 and NME2 engage in mutual functions in the nucleus, possibly performing specific functions in their homomeric states. Finally, we demonstrated that fluorophores fused to the N-termini of NME polypeptides produce the largest FRET effect and thus recommend this orientation for use in similar studies.

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Techniques for detecting protein-protein interactions in living cells: principles, limitations, and recent progress

Cited by 57 publications

References 98 publications

At the intersection of exocytosis and endocytosis in plants

At the intersection of exocytosis and endocytosis in plants

Systemin‐mediated long‐distance systemic defense responses

The Subcellular Localization and Oligomerization Preferences of NME1/NME2 upon Radiation-Induced DNA Damage

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