Conformational changes at protein–protein interaction followed online with an SAW biosensor

Klumpers, Fabian; Götz, Ulrich; Kurtz, Tanja; Herrmann, Christian; Gronewold, Thomas M. A.

doi:10.1016/j.snb.2014.07.064

Cited by 10 publications

(5 citation statements)

References 15 publications

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“…Exactly how Ras facilitates the interaction between NORE1A and PP1A and Rb is not entirely clear. However, binding of Ras to NORE1A induces a conformational change (46), and this may promote the interaction between NORE1A and its binding partners.…”

Section: Nore1a Forms An Endogenous Ras-regulated Complex Withmentioning

confidence: 99%

Ras Regulates Rb via NORE1A

Barnoud

Donninger²,

Clark

2016

Journal of Biological Chemistry

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Mutations in the Ras oncogene are one of the most frequent events in human cancer. Although Ras regulates numerous growth-promoting pathways to drive transformation, it can paradoxically promote an irreversible cell cycle arrest known as oncogene-induced senescence. Although senescence has clearly been implicated as a major defense mechanism against tumorigenesis, the mechanisms by which Ras can promote such a senescent phenotype remain poorly defined. We have shown recently that the Ras death effector NORE1A plays a critical role in promoting Ras-induced senescence and connects Ras to the regulation of the p53 tumor suppressor. We now show that NORE1A also connects Ras to the regulation of a second major prosenescent tumor suppressor, the retinoblastoma (Rb) protein. We show that Ras induces the formation of a complex between NORE1A and the phosphatase PP1A, promoting the activation of the Rb tumor suppressor by dephosphorylation. Furthermore, suppression of Rb reduces NORE1A senescence activity. These results, together with our previous findings, suggest that NORE1A acts as a critical tumor suppressor node, linking Ras to both the p53 and the Rb pathways to drive senescence.Ras mutations are the most frequent oncogenic events in human cancer and can be found in ϳ30% of all human cancers (1). In experimental systems, activated forms of Ras can be powerfully transforming, and transgenic animal models have validated the role of Ras activation in tumorigenesis (2, 3). However, despite the extensive evidence linking Ras to transformation and tumor development, activated Ras can also promote a state of irreversible cell cycle arrest called oncogeneinduced senescence (4, 5). This tendency for deregulated Ras activity to provoke senescence can be observed in Ras-driven tumors (6). It appears that senescence provides a potent barrier to suppress the development of Ras-driven cancer because malignant tumors lose the senescence phenotype (6). The exact mechanisms by which Ras can promote senescence are not completely understood, but it appears that the main Ras senescence pathways involve the p53 and retinoblastoma (Rb) 2 tumor suppressors (7). Initial evidence in mouse embryonic fibroblasts (MEFs) suggests that loss of functional p53 or Rb pathways alone is sufficient for Ras to bypass senescence (5). More recent studies have shown that, in vivo, suppression of p53 function (8) or Rb (9) enhances Ras-mediated transformation in murine systems. However, human systems may require inactivation of both p53 and Rb for full senescence evasion (10). Therefore, inactivation of the p53 and Rb senescence pathways may be essential for Ras-induced transformation.In addition to the classic trio of growth-promoting Ras effector proteins, Raf, PI3K, and RalGDS, Ras also interacts with growth-suppressing effector proteins, including NORE1A (RASSF5) (11,12). NORE1A is a member of the RASSF family of tumor suppressors that is frequently down-regulated during tumor development, and its inactivation has been linked to a rare familial cancer...

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Section: Nore1a Forms An Endogenous Ras-regulated Complex Withmentioning

confidence: 99%

Ras Regulates Rb via NORE1A

Barnoud

Donninger²,

Clark

2016

Journal of Biological Chemistry

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“…The increase in mass on the biosensor surface upon drug binding induces a phase shift of the acoustic wave, whereas changes in the viscoelasticity on the surface -e.g. by a conformational change of the immobilized protein -mainly affect its amplitude [9,10]. While kinetic-binding analysis of small molecules by SPR is nowadays performed in a routine fashion for soluble protein targets like proteases or kinases, it is still challenging for integral membrane proteins like GPCRs.…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

Label-free detection of small-molecule binding to a GPCR in the membrane environment

Heym

Hornberger

Lakics

et al. 2015

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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“…36° Y-cut LiTaO 3 , 41° Y-cut LiNbO 3 , etc) [9][10][11][12]. In both modalities, SH-SAWs are strongly confined to the surface thus determining an enhanced sensitivity towards mass loading as well as changes of viscosity, viscoelasticity and dielectric permittivity near the surface [13,14].…”

Section: Introductionmentioning

confidence: 99%

A high-performance lab-on-a-chip liquid sensor employing surface acoustic wave resonance

Kustanovich¹,

Yantchev²,

Kirejev³

et al. 2017

J. Micromech. Microeng.

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We demonstrate herein a new concept for lab-on-a-chip in-liquid sensing, through integration of surface acoustic wave resonance (SAR) in a one-port configuration with a soft polymer microfluidic delivery system. In this concept, the reflective gratings of a one-port surface acoustic wave (SAW) resonator are employed as mass loading-sensing elements, while the SAW transducer is protected from the measurement environment. We describe the design, fabrication, implementation, and characterization using liquid medium. The sensor operates at a frequency of 185 MHz and has demonstrated a comparable sensitivity to other SAW in-liquid sensors, while offering quality factor (Q) value in water of about 250, low impedance and fairly low susceptibility to viscous damping. For proof of principle, sensing performance was evaluated by means of binding 40 nm neutravidin-coated SiO2 nanoparticles to a biotin-labeled lipid bilayer deposited over the reflectors. Frequency shifts were determined for every step of the affinity assay. Demonstration of this integrated technology highlights the potential of SAR technology for in-liquid sensing.

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Conformational changes at protein–protein interaction followed online with an SAW biosensor

Cited by 10 publications

References 15 publications

Ras Regulates Rb via NORE1A

Ras Regulates Rb via NORE1A

Label-free detection of small-molecule binding to a GPCR in the membrane environment

A high-performance lab-on-a-chip liquid sensor employing surface acoustic wave resonance

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