biochemistry

Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes

In the last decades enormous advances have been made in characterizing the atomic and molecular structure of respiratory chain supercomplexes. *

However, it still remains a challenge to stitch this refined spatial atomistic description with functional information provided by biochemical studies of isolated protein material. Development of functional assays that detect respiratory chain complexes in their native membrane environment contribute to address the open questions related to the role played by their association and interactions. *

In the article “Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes” Daniel G. Cava, Julia Alvarez-Malmagro, Paolo Natale, Sandra López-Calcerrada, Iván López-Montero, Cristina Ugalde, Jose Maria Abad, Marcos Pita, Antonio L. De Lacey and Marisela Vélez present a characterization assay in which a functionalized gold electrode is modified with mitochondrial membrane fragments that allows monitoring electrochemically the activity of different respiratory chain complexes immersed in the mitochondrial membrane. *

Daniel G. Cava et al. measure the intensity of the reducing current of the electron mediator CoQ1 at the electrode surface and its variation upon addition of the corresponding enzymatic substrates. The activities of Complex I, Complex II and Complex III were monitored by the way in which they reduce the current, reflecting the amount of quinone reduced by the complexes in the presence of their substrates. *

The authors detect that CoQ1H2 produced by Complex I remains partially trapped within the membrane and is more easily oxidized by Complex III or the electrode than the quinone reduced by Complex II. *

Atomic Force Microscopy (AFM) was used to image the topography of the membrane modified electrode. NanoWorld Pyrex-Nitride Silicon-Nitride AFM probes (PNP-DB, diving board shaped cantilevers, the short AFM cantilever with a typical force constant of 0.48 N/m and 67 kHz resonance frequency) were used. *

The surfaces analysed were the electrodes. The two surfaces imaged are the same previously polished electrodes used for electrochemical measurements. The microscope sample holder was adapted in-home to support the electrodes. Two surfaces were analysed: the polished gold functionalized with 4-aminothiophenol and the electrode after incubation with mitochondria subparticles prepared similarly to the electrodes used for the electrochemical measurements.*

Fig. 2 from Daniel G. Cava et al 2024 “Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes”
QCM and AFM characterization of modified gold. Panel A shows the frequency (left, black) and dissipation (right red) changes detected on a gold covered quartz crystal previously modified with a 4-ATP after injection in the chamber of the mitochondrial fragments at the time point indicated by the thick arrow. Panel B show AFM images of the surface topography of a modified gold electrode before (left) and after (right)incubation with the mitochondrial membrane. The inset below shows the height profile of the lines indicated in the images.

*Daniel G. Cava, Julia Alvarez-Malmagro, Paolo Natale, Sandra López-Calcerrada, Iván López-Montero, Cristina Ugalde, Jose Maria Abad, Marcos Pita, Antonio L. De Lacey and Marisela Vélez
Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes
Electrochimica Acta, Volume 484, 20 April 2024, 144042
DOI: https://doi.org/10.1016/j.electacta.2024.144042

The article “Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes” by Daniel G. Cava, Julia Alvarez-Malmagro, Paolo Natale, Sandra López-Calcerrada, Iván López-Montero, Cristina Ugalde, Jose Maria Abad, Marcos Pita, Antonio L. De Lacey and Marisela Vélez is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third-party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.

Actin filaments on Mica with APTES

Courtesy of Prof. Noriyuki Kodera, nanoLSI, Kanazawa University, Japan we could upload two new images and a very nice High Speed Atomic Force Microscopy (HS-AFM) video of Actin filaments on Mica with APTES to the image gallery and the video gallery on https://www.highspeedscanning.com/.

NanoWorld USC-F1.2-k0.15 Ultra-Short AFM cantilevers [C = 0.15 N/m; fo = 1200 kHz] were used for the high speed imaging.

HS-AFM Images of actin filaments on Mica with APTES. Buffer:100 mM KCl, 2 mM MgCl2, 1 mM EGTA, 20 mM Imidazole-HCl, pH7.6 (a) 250×250 nm2 and (b) 400×400 nm2. Image courtesy of Prof. Kodera, nanoLSI, Kanazawa University, Japan. NanoWorld USC-F1.2-k0.15 AFM probes from the Ultra-Short Cantilever (USC) series were used for the High Speed Atomic Force Microscopy — HS-AFM Images of actin filaments on Mica with APTES. Buffer:100 mM KCl, 2 mM MgCl2, 1 mM EGTA, 20 mM Imidazole-HCl, pH7.6 (a) 250×250 nm2 and (b) 400×400 nm2. Images courtesy of Prof. Kodera, nanoLSI, Kanazawa University, Japan. NanoWorld USC-F1.2-k0.15 AFM probes were used for the High Speed Atomic Force Microscopy

The HighSpeedScanning webpage is dedicated to presenting information about ultra high frequency AFM probe solutions for High Speed AFM ranging from already commercialized AFM probes such as the Arrow^TM UHF and NanoWorld Ultra-Short Cantilever (USC) series to AFM probes that are still under development.

Additionally to the High-Speed AFM images and videos researchers worldwide kindly have provided us with so that we can share them with the whole High Speed AFM community you will also find a list of links and references dedicated to High-Speed AFM on https://www.highspeedscanning.com/high-speed-scanning.html

We are aware that these lists are far from complete and we are constantly working on keepting them up to date. If your research institute or company is working with High Speed AFM (HS-AFM) using our AFM probes or if you have published articles with images that were achieved with our High Speed AFM probes annd you find that are missing from our list then please let us know via email: info@highspeedscanning.com if you would like us to add them to the list of references .

Tissue mechanics and expression of TROP2 in oral squamous cell carcinoma with varying differentiation

Atomic Force Microscopy ( AFM ) can be utilized to determine the mechanical properties of tumor tissues in different kinds of cancers, for example breast cancer, liver cancer and lung cancer.

Oral squamous cell carcinoma (OSCC) is a common subtype of head and neck and other malignant tumors that occurs in increasing numbers. It is therefore important to learn more about the biological factors connected with the early diagnosis and treatment of OSCC. *

The human trophoblast cell surface antigen 2 (TROP2), which is also called tumor-associated calcium signal transduction-2 (TACSTD-2), is a surface glycoprotein encoded by TACSTD. *

Among the various biochemical mechanisms involved in tumorigenesis, the role of β-catenin has been studied extensively. This has shed light on the biological functions of TROP2 and its use as a prognostic biomarker for OSCC. *

TROP2 regulates tumorigenic properties including cancer cell adhesion, invasion, and migration and is overexpressed in many human cancers. Inhibiting TROP2 expression has shown promise in clinical applications. *

In the article “Tissue mechanics and expression of TROP2 in oral squamous cell carcinoma with varying differentiation” Baoping Zhang, Shuting Gao, Ruiping Li, Yiting Li, Rui Cao, Jingyang Cheng, Yumeng Guo, Errui Wang, Ying Huang and Kailiang Zhang investigate the role of TROP2 in OSCC patients using a combination of biophysical approaches including atomic force microscopy. *

The authors demonstrate the tissue morphology and mechanics of OSCC samples during tumor development using NanoWorld Pointprobe® CONTR AFM probes for the Atomic Force Microscopy described in the article and they believe that their findings will help develop TROP2 in accurately diagnosing OSCC in tumors with different grades of differentiation. *

Figure 5 from Baoping Zhang et al. “Tissue mechanics and expression of TROP2 in oral squamous cell carcinoma with varying differentiation”:
Surface morphology of OSCC tissue sections via AFM detection, irregular morphology appeared in the low differentiation
NanoWorld Pointprobe CONTR AFM probes were used for the Atomic Force Microscopy — Figure 5 from Baoping Zhang et al. “*Tissue mechanics and expression of TROP2 in oral squamous cell carcinoma with varying differentiation*”:
Surface morphology of OSCC tissue sections via AFM detection, irregular morphology appeared in the low differentiation

*Baoping Zhang, Shuting Gao, Ruiping Li, Yiting Li, Rui Cao, Jingyang Cheng, Yumeng Guo, Errui Wang, Ying Huang and Kailiang Zhang
Tissue mechanics and expression of TROP2 in oral squamous cell carcinoma with varying differentiation
BMC Cancer volume 20, Article number: 815 (2020)
DOI: https://doi.org/10.1186/s12885-020-07257-7

Please follow this external link to read the whole article: https://rdcu.be/cfC9G

Open Access : The article “Tissue mechanics and expression of TROP2 in oral squamous cell carcinoma with varying differentiation” by Baoping Zhang, Shuting Gao, Ruiping Li, Yiting Li, Rui Cao, Jingyang Cheng, Yumeng Guo, Errui Wang, Ying Huang and Kailiang Zhang is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.