新的分子間相互作用分析儀微量熱泳動(dòng)儀---MST 用戶評(píng)價(jià)舉例

Our recent research is focused on the molecular basis of herbicide resistance and exploiting new target sites of herbicide, such as PPI of AHAS. Via MST, we measured the Protein-Protein interaction, that have been difficult to detect and quantify with ITC because of instability of the protein and very little heat changes of our samples. The MST technology can be used as a complementary technology to the traditional methods in these fields.

We can strongly recommend the MST technology for its low material consumption, short measurement times and broad application range in the field of molecule interaction studies.

我們目前致力于除草劑抗性的分子機(jī)制以及尋找新的除草劑靶點(diǎn)（比如AHAS亞基間相互作用的界面）等方面的研究。通過(guò)MST，我們測(cè)量了蛋白質(zhì)-蛋白質(zhì)相互作用，而我們的樣品使用ITC方法測(cè)定沒(méi)有得到結(jié)果，因?yàn)槲覀兊臉悠贩�(wěn)定性較差并且相互作用熱量變化極小。所以在這些研究領(lǐng)域，MST技術(shù)能夠很好地與傳統(tǒng)的技術(shù)互補(bǔ)。

王大成

Our resent research is focused on the critical pathogenic protein & complex from pathogenic microbes and The Protein structure-functional basis for some endogenous disease. Via MST, we measured the Protein-DNA interaction, that have been difficult to detect and quantify with other standard biomolecule interaction technologies (like SPR and ITC) because of the conformation/immobilization-sensitive and very little heat changes of our samples. MST technology can be used as a complementary technology to the traditional methods in these fields.

We can strongly recommend MST technology for its low material consumption, short measurement times and broad application range for molecule-interaction studies.

我們非常推薦MST技術(shù)的一些特點(diǎn)，如低樣品使用量、很短的測(cè)量時(shí)間以及在分子間相互作用方面很廣泛的應(yīng)用。

谷立川

Part of our research focus on the protein structural-function basis for iron metabolism of pathogenic microorganisms. We measured the dissociated constant for periplasmic binding protein and siderophore through MST technology. Because siderophore is extremely insoluble, it's difficult for us to complete that task via other methods such as ITC and SPR. We strongly recommend MST technology for its low material consumption, short measurement time and very wide application range.

我們目前一部分課題是關(guān)于致病微生物鐵代謝相關(guān)蛋白的結(jié)構(gòu)與功能。通過(guò)MST技術(shù)，我們測(cè)量了周質(zhì)空間轉(zhuǎn)鐵蛋白與鐵載體的解離常數(shù)，我們的樣品很難通過(guò)其他傳統(tǒng)方法（例如SPR和ITC）來(lái)測(cè)量因?yàn)樾》肿拥娜芙舛葮O低，MST技術(shù)的我們得到了理想的結(jié)果。我們非常推薦MST技術(shù)的一些特點(diǎn)，如極低的樣品使用量，較快的測(cè)量時(shí)間，以及十分廣泛的應(yīng)用。

“We routinely assess interaction affinity for both small molecule and biologics projects, with NanoTemper Technologies’ Microscale Thermophoresis being the most recent addition to the pool of instruments we use to carry out these measurements. It has proved a valuable tool for characterising small molecule-protein and protein-protein interactions, as well as for the study of protein aggregation concentration determination. There is very good agreement with other technologies such as Surface Plasmon Resonance (SPR) and Isothermal Titration Calorimetry (ITC), and we are particularly appreciative of this new technology because of the extremely low protein consumption and relatively short time required for the assay setup. NanoTemper customer support has been a key factor in enabling us to familiarise ourselves with the new technology. We would like to deploy increasing numbers of applications based on MST technologies and continue to interact with NanoTemper Technologies Company, a dynamic, scientifically driven company.”

（編者譯）

我們經(jīng)常需要測(cè)量小分子和生物制劑的相互作用親和力，我們最近使用的最新一個(gè)設(shè)備就是NanoTemper的微量熱泳動(dòng)技術(shù)。結(jié)果證明了MST是一個(gè)有價(jià)值的工具，用于測(cè)量小分子-蛋白質(zhì)、蛋白質(zhì)之間的相互作用，以及判斷蛋白質(zhì)形成聚集體的濃度。MST技術(shù)與傳統(tǒng)的其他技術(shù)如表面等離子體共振(SPR)和等溫滴定量熱法(ITC)的結(jié)果完全吻合，我們特別喜歡這個(gè)新技術(shù)的一些特點(diǎn)：極低的蛋白樣品用量和很短的實(shí)驗(yàn)設(shè)置時(shí)間。NanoTemper的技術(shù)支持非常及時(shí)有效，使我們能夠很快熟悉這個(gè)新技術(shù)。我們很愿意使用越來(lái)越多MST儀器并繼續(xù)與NanoTemper技術(shù)公司相互合作，因?yàn)檫@個(gè)一個(gè)積極向上的、技術(shù)先進(jìn)的公司。

 

（編者譯）

我們的主要工作是根據(jù)結(jié)構(gòu)設(shè)計(jì)藥物。比如，細(xì)菌tRNA修飾酶-tRNA鳥嘌呤轉(zhuǎn)糖苷酶(Tgt)與小分子抑制劑以及tRNA的相互作用。雖然我們無(wú)法直接衡量這個(gè)酶的與小分子鳥嘌呤以及7-deaza-7-aminomethyl的相互作用，但是由于Tgt結(jié)合大分子tRNA是有很明顯的熱泳動(dòng)效應(yīng)變化，我們可以通過(guò)MST來(lái)測(cè)量各自的Kd值。此外，MST測(cè)量出來(lái)lin-benzoguanine-based MST Tgt抑制劑(300 - 500 Da) 的Kd值極好地吻合先前確定通過(guò)酶動(dòng)力學(xué)研究得到的Ki值。因此，這個(gè)快速和易于使用的方法提供了一個(gè)非常好的方法用于替代放射性酶活性測(cè)定，而后者是我們之前一直用來(lái)測(cè)量Tgt抑制劑親和力的方法。我們使用到目前為止找出綁定的親和力Tgt抑制劑。此外,我們還研究了一個(gè)志賀氏桿菌III型分泌系統(tǒng)分子伴侶蛋白與其結(jié)合物（合成的多肽）的相互作用。通過(guò)MST，我們能通過(guò)極低量的蛋白質(zhì)來(lái)測(cè)定Kd值，而且其結(jié)果幾乎符合傳統(tǒng)的等溫滴定量熱法ITC等技術(shù)所得到的數(shù)據(jù)。

We use systems biology to study transcription regulatory phenomena on a genome-wide scale in the context of host-pathogen interactions and cancer.

Microscale Thermophoresis (MST) thereby has been extremely useful to rapidly quantify relevant protein-nucleic acid and protein-protein interactions directly in cellular lysates without going through the hassles of purification.

In my group we explore the structural, biophysical, and cellular outcomes of protein complex formation at membrane-bound receptors.

We use Microscale Thermophoresis (MST) in addition to other methods including isothermal titration calorimetry and surface plasmon resonance and find a good agreement between the methods. We are very pleased with the low material consumption, short measurement times and broad application range of MST.

 

We study the structural and molecular biology of bacterial adhesins and cell-surface filaments with respect to their function in bacterial pathogenesis, with the ultimate aim of developing a new generation of virulence-targeted antimicrobials.

MST really is opening up the easy determination of some Kd's that were hard to get to with other methods. So far, we measured protein-protein, protein-glycan, protein-small compound and protein-cofactor interactions with the Monolith NT.115.”

 

We are studying the molecular mechanisms of signal transduction processes involving Rho GTPases and are developing small molecule inhibitors and other strategies that interfere with specific Rho protein functions in leukemia, lymphoma and lung cancer.

We are very enthusiastic about MST as it quickly enabled us to measure protein:protein and protein:small molecule interactions that have been difficult to detect and quantify with standard biomolecule interaction technologies in the past years. MST provides us with a unique tool to validate the lead inhibitors that bind to specific sites of Rho GTPases or their regulators/effectors .

（編者譯）

我們正在研究Rho GTPases的信號(hào)轉(zhuǎn)導(dǎo)的分子機(jī)制的，并開發(fā)小分子抑制劑和其他方法用于特異性地干擾白血病、淋巴瘤、肺癌中的Rho蛋白質(zhì)功能。

我們很喜歡MST技術(shù)，因?yàn)樵谶^(guò)去的幾年里MST幫助我們快速完成測(cè)量蛋白質(zhì)-蛋白質(zhì)和蛋白質(zhì)-小分子間的相互作用，很多時(shí)候很難使用其他標(biāo)準(zhǔn)的生物分子互作技術(shù)完成。MST為我們提供了一個(gè)獨(dú)特的工具來(lái)驗(yàn)證重要的抑制劑是否結(jié)合到Rho GTPases的特異性位點(diǎn)或者結(jié)合到他的調(diào)節(jié)物/效應(yīng)物。

 

“Our lab is developing diagnostic tools based on protein biochips and is studying the basic mechanisms underlying herpes virus infections” We are using MST for elucidating the function of viral proteins and its interaction with other viral and host cell proteins.

MST is simple to use and was quickly established in our lab. In particular we do appreciate the low instrument, consumables and maintenance costs associated with this new technology”

“We are interested in understanding the interactions between cells and organisms by investigating the role of polysaccharides exposed on cell surfaces and secreted polysaccharide signal molecules. We are applying MST to determine structural requirements for recognition of complex polysaccharides and the role of ligand-receptor interactions in the relationships between different cells and organisms.

MST is very useful for my lab since it allows to measure interactions in solution even in complex samples of membrane proteins. Also the small amount of sample material needed and the broad range of applications are very advantageous.”

（編者譯）
我們的實(shí)驗(yàn)室通過(guò)研究暴露在細(xì)胞表面的多糖和分泌型多糖信號(hào)分子來(lái)揭示細(xì)胞與細(xì)胞以及有機(jī)體的相互作用。我們使用MST來(lái)測(cè)量復(fù)雜的多糖的結(jié)合以及配體-受體的結(jié)合以闡明不同細(xì)胞或機(jī)體間的相互作用。

My lab focuses on the elucidation of the structure and function of membrane proteins. This topic includes the investigation of protein-protein and protein-substrate interactions. We are employing MST in binding studies of membrane sensors to their transducers as well as soluble transcription factors to DNA. We also plan for investigations of receptor – ligand interactions with this exciting new technique. MST turned out to be an extremely useful method in our lab to determine binding constants. It is advantageous that the fast and handy.

（編者譯）

我們的實(shí)驗(yàn)室專注于研究膜蛋白的結(jié)構(gòu)和功能，包括蛋白-蛋白以及蛋白-底物相互作用的研究。我們使用MST來(lái)研究膜蛋白感應(yīng)器分子-傳感器分子-轉(zhuǎn)錄因子-DNA的結(jié)合反應(yīng)。MST顯示出極好的優(yōu)點(diǎn)，包括快速和簡(jiǎn)單測(cè)量，樣品消耗量很少等等。

Our lab focuses on the mechanisms and organization of DNA packaging inside the cell nucleus. We study the biochemical properties of molecular machines, the chromatin remodelers that regulate DNA accessibility and switch between ‘on’ and ‘off’ states of genes.

We are using MST for studying the activity of chromatin remodeling enzymes and to quantify their affinities towards DNA, RNA and other proteins.

MST is a big step forward towards quantitative biochemistry. MST is simple to use and was quickly established in our lab. It is a good alternative to the traditional electrophoretic mobility shift assays.

（編者譯）

我們的實(shí)驗(yàn)室專注于研究細(xì)胞和之中的DNA包裝的機(jī)制，研究分子機(jī)器的生化特性，染色質(zhì)重塑調(diào)節(jié)DNA組裝和調(diào)節(jié)基因的開關(guān)狀態(tài)。我們通過(guò)MST來(lái)研究染色質(zhì)重組化酶并且定量他們與DNA，RNA以及其他蛋白之間的親和力。MST將定量生化技術(shù)推向一個(gè)新的臺(tái)階因?yàn)槠浜?jiǎn)單易學(xué)，并且非常好的取代了傳統(tǒng)的EMSA技術(shù)。

We are interested in the basic mechanisms in epigenetics which define and maintain the histone code and are using MST to measure the binding of “reader”-proteins to modified histone peptides as well as the activity and inhibition of “writer” enzymes including kinases, demethylases and methylases.

The new MST technology is easy to use and a good alternative to our standard enzymatic assays which are radioactive and measure endpoints only. It allows us to easily measure affinities for protein-peptide interactions in solution.

（編者譯）

我們對(duì)實(shí)驗(yàn)胚胎學(xué)的定義和維護(hù)組蛋白準(zhǔn)則的基本機(jī)制很感興趣，并且使用MST來(lái)測(cè)量“Reader”蛋白和修飾后的組蛋白多肽的結(jié)合以及“writer”酶（包括激酶、去甲基化酶、甲基化酶）的激活劑和抑制劑作用。

The work of our group gave rise to the novel concept that under certain conditions matrix components may act as endogenous “danger” signals, which are recognized by innate immunity receptors, and are capable to trigger an inflammatory response reaction.

We are using MST to measure the interactions of endogenous and in vitro mutagenized proteoglycans of the extracellular matrix with their receptors and are very satisfied with the results obtained so far. The affinities are consistent with the biological readouts and confirm previous results gained with immunprecipitation and binding assays.

MST is a new technology we can definitely recommend for obtaining robust quantitative affinity data.”

（編者譯）

我們發(fā)現(xiàn)在特定化井下某些基質(zhì)成分會(huì)變成內(nèi)源性的“危險(xiǎn)”信號(hào)，而這些異常的成分會(huì)被先天免疫受體識(shí)別，并且能夠引發(fā)炎癥反應(yīng)。

我們使用MST來(lái)測(cè)量細(xì)胞外基質(zhì)中的內(nèi)源物/體內(nèi)突變的蛋白聚糖與他們相關(guān)受體的反應(yīng)，并且非常滿意目前獲得的所有數(shù)據(jù)，所得到的數(shù)據(jù)與其他生物方法預(yù)測(cè)的完全吻合并且驗(yàn)證了之前使用免疫沉淀和結(jié)合試驗(yàn)所獲得的數(shù)據(jù)。

MST是一種非常新的技術(shù)，我們推薦這個(gè)技術(shù)可以用來(lái)獲得非常好的定量的親和力數(shù)據(jù)。

We are studying the processing of microRNA precursors In Drosophila and are using MST to measure the binding of nucleolytic Enzymes and their specificity factors to RNA substrates.

The small amount of protein sample needed and the much faster measuring time compared with e.g. gel-shift assays are particular strengths of this new technique. It enables us to ask questions that we could not address before.

（編者譯）

我們正在研究果蠅前體細(xì)胞的MicroRNA發(fā)生過(guò)程，并且使用MST來(lái)測(cè)量核酶即特異性的因子結(jié)合到RNA上的反應(yīng)。MST極少的蛋白樣品的消耗量和極短的測(cè)量時(shí)間使得他成為一個(gè)非常有價(jià)值的技術(shù)，它能幫助我們回答很多之前無(wú)法解釋的問(wèn)題。

Membrane interaction and fusion plays a fundamental role in many cellular processes such as intracellular trafficking, fertilization, tissue formation or viral infection.

With the new MST technology we were able to measure the interaction of membrane vesicles, mediated by coiled coil-forming peptides. The technology requires only little sample material and is enabling for this type of experiments”（編者譯）

生物膜的融合以相互作用在細(xì)胞的多項(xiàng)過(guò)程中（細(xì)胞內(nèi)運(yùn)輸，受精，組織形成以及病毒感染）起到非常重要的基礎(chǔ)作用。通過(guò)MST技術(shù)，我們能夠測(cè)量螺旋狀多肽介導(dǎo)的囊泡的相互作用。該技術(shù)僅僅需要極少的樣品材料即可完成測(cè)量。

We are interested in the biophysical properties of soluble and membrane bound biomolecules and are using MST to measure binding affinities and complex formation.

MST is straightforward and fits well in our technology portfolio to cover a broad spectrum of synthetic biomolecules including proteins, peptides and nucleic acids.

（編者譯）

生物膜的融合以相互作用在細(xì)胞的多項(xiàng)過(guò)程中（細(xì)胞內(nèi)運(yùn)輸，受精，組織形成以及病毒感染）起到非常重要的基礎(chǔ)作用。通過(guò)MST技術(shù)，我們能夠測(cè)量螺旋狀多肽介導(dǎo)的囊泡的相互作用。該技術(shù)僅僅需要極少的樣品材料即可完成測(cè)量。我們對(duì)游離的和膜上生物分子的生物物理學(xué)性質(zhì)感興趣，并且使用MST來(lái)測(cè)量結(jié)合親和力和復(fù)合物的構(gòu)象。

 

 

We are interested in molecular architectures especially to stabilize and analyze challenging smell receptors that belong to G protein-coupled receptors (GPCRs). GPCR is a super-family of receptors linked to many diseases. We want to measure the interactions of smell receptors with odorant that are very low molecular weight ligands. In the past that has been extremely difficult to detect and quantify other common molecular interaction technologies including SPR, and nearly impossible to use ITC for odorant-binding measurement because of the conformation and immobilization-sensitivities, or difficult and costly to obtain sufficient amount of receptors. The MST technology is a superb technology, it not only is label-free, hence reducing the measuring artifacts, but it also is surface-free that provides true measurement of finest molecular interactions. Thus it allows us much better analysis of the GPCR with their small-molecule ligands. We strongly recommend MST technology for its low material consumption (a few micrograms in a few microliters), short measurement times, inexpensive consumables, and broad application range in all molecule-interaction study.