Name: airid-based proximity labeling for protein-protein interaction in plants
Uploaded: 2022-09-16T14:20:00
Description: Watch this Scientific Journal Video about AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants at JoVE.com

This work was supported by the National Natural Science Foundation of China (Grant No. 32000197 to X.H.), the Special Financial Grant from the China Postdoctoral Science Foundation (Grant No. 2019T120467 to X.H.)

1. Preparation of plant material
<ol>
	<li>Cucumis sativus (Cucumber) is employed for experimental analysis. Put the seeds in water, incubate at 50 &#176;C for 20 min, and then place the seeds on filter paper on a Petri plate for 12-16 h.</li>
	<li>Afterward, transfer the seeds to pots containing soil (purchased commercially) and grow in a climatic chamber at 23 &#176;C temperature and 16 h light and 8 h dark photoperiod.</li>
	<li>Maintain the plants in the climate chamber for 3-4 weeks until...

<ol>
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(159), e60728 (2020).</li><li>Kido, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AirID,+a+novel+proximity+biotinylation+enzyme,+for+analysis+of+protein-protein+interactions.">AirID, a novel proximity biotinylation enzyme, for analysis of protein-protein interactions.</a> eLife. 9, 54983 (2020).</li><li>Hu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overexpressing+7-hydroxymethyl+chlorophyll+a+reductase+alleviates+non-programmed+cell+death+during+dark-induced+senescence+in+intact+Arabidopsis+plants.">Overexpressing 7-hydroxymethyl chlorophyll a reductase alleviates non-programmed cell death during dark-induced senescence in intact Arabidopsis plants.</a> Biomolecules. 11 (8), 1143 (2021).</li><li>Ammiraju, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Oryza+bacterial+artificial+chromosome+library+resource:+construction+and+analysis+of+12+deep-coverage+large-insert+BAC+libraries+that+represent+the+10+genome+types+of+the+genus+Oryza.">The Oryza bacterial artificial chromosome library resource: construction and analysis of 12 deep-coverage large-insert BAC libraries that represent the 10 genome types of the genus Oryza.</a> Genome Research. 16 (1), 140-147 (2006).</li><li>Liu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Albino+Leaf+2+is+involved+in+the+splicing+of+chloroplast+group+I+and+II+introns+in+rice.">Albino Leaf 2 is involved in the splicing of chloroplast group I and II introns in rice.</a> Journal of Experimental Botany. 67 (18), 5339-5347 (2016).</li><li>Branon, T. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+proximity+labeling+in+living+cells+and+organisms+with+TurboID.">Efficient proximity labeling in living cells and organisms with TurboID.</a> Nature Biotechnology. 36 (9), 880-887 (2018).</li><li>Nakano, S., Niwa, M., Asano, Y., Ito, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Following+the+evolutionary+track+of+a+highly+specific+l-Arginine+oxidase+by+reconstruction+and+biochemical+analysis+of+ancestral+and+native+enzymes.">Following the evolutionary track of a highly specific l-Arginine oxidase by reconstruction and biochemical analysis of ancestral and native enzymes.</a> Applied and Environmental Microbiology. 85 (12), 00459 (2019).</li><li>Gingras, A. -. C., Abe, K. 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In the current experiment, AirID was used for proximity labeling, which Kido et al. developed through an algorithm of ancestral enzyme reconstruction using a large genome dataset and five conventional BirA enzymes5. Random mutations were used in traditional evolutionary protein engineering to enhance activity9,10 as random mutations cannot produce dynamic sequence changes. Compared to other PL enzymes, AirID has several advantages. Previou...

Various cellular proteins work under the biologically regulatory system, and protein-protein interactions (PPIs) are a part of this system and the basis of many cellular processes. Besides PPIs, the function of natural proteins is post-translationally promoted via various modifications such as the formation of complex, ubiquitination, and phosphorylation. Therefore, studying PPIs is significant to understanding the possible function of target proteins. PPIs have been carried out using various technologies such as mass spectrometry analysis after immunoprecipitation (IP-MS analysis)1, yeast two-hybrid system (Y2H)2, also cell-free based arrays3. These methods explored various vital findings in the field of research. However, these methods have some drawbacks; for example, Y2H is a time-consuming, expensive strategy that necessitates building the target species&#39; Y2H library.
Additionally, the Y2H technique uses yeast, a heterologous single-cell eukaryotic organism, which could not accurately reflect the cellular state of higher eukaryotic cells. The IP-MS is unsuitable for high hydrophobicity proteins and shows low efficiency in capturing weak PPIs. Various essential proteins in plants such as nucleotide-binding domain and leucine-rich repeat-containing (NLR) proteins and receptor-like kinases (RLKs) are expressed at a low level and mostly interact with other proteins transiently; therefore, using these methods is insufficient for understanding the mechanisms underlying the regulation of these proteins3.
A new technique called proximity biotinylation (PB) helps researchers identify PPIs. PB depends on PL enzymes, which attach to the protein of interest (POI), and when partner protein comes near POI, the PL attaches a chemical biotin tag to the partner protein. Further, the tagged protein can be identified and can quickly know which partner protein attaches to the target protein5. Previous studies proved that BioID and TurboID are successful tools for PPIs, especially in plants, but they have certain limitations4. BioID needs a high level of biotin for labeling partner proteins, which takes more than 16 h. Compared to BioID, the TurboID is more beneficial as it labels protein in 10 min and can label the partner protein at room temperature (RT). It is also toxic to cells in certain conditions and tags those proteins that do not show interaction with the protein of interest.
To overcome these issues, AirID, developed by Kido et al., is more efficient than the rest of the labeling enzymes, although the sequence similarity is 82% between BioID and AirID5. To check the efficiency of AirID, we conducted an experiment by using a POI with known associates. This experiment confirmed that AirID could undoubtedly label associated proteins in plant cells. AirID is a valuable enzyme for analyzing PPIs in vitro and in cells. It creates less toxicity and is less erroneous in time taken processes than TurboID to tag non-partners, leading to killing the cell. It demonstrates that AirID is more competitive than other labeling enzymes for proximity biotinylation. It is more accurate, has more potential in time-taking processes, and less toxic in vitro and in living cells. The current protocol describes the identification of interacting proteins of APRR2 using AirID as a PL enzyme; furthermore, the method can be applied to other proteins to investigate PPIs in plant species.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Acetosyringone</td>
			<td>Beijing solaribo science and technology Co.Ltd</td>
			<td>S1519</td>
			<td></td>
		</tr>
		<tr>
			<td>Acryl/Bis 30% solution</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>1510KA4528</td>
			<td></td>
		</tr>
		<tr>
			<td>Agar</td>
			<td>BioFroxx GmbH</td>
			<td>D64683</td>
			<td></td>
		</tr>
		<tr>
			<td>Agarose</td>
			<td>tsingke (Shanghai) Co.Ltd</td>
			<td>TSJ001</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium bicarbonate</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>G313BA0018</td>
			<td></td>
		</tr>
		<tr>
			<td>Biotin</td>
			<td>BBI life Sciences</td>
			<td>G908BA0012</td>
			<td></td>
		</tr>
		<tr>
			<td>CaCl2</td>
			<td>BBI life Sciences</td>
			<td>E209BA0008</td>
			<td></td>
		</tr>
		<tr>
			<td>Competent cells GV3101</td>
			<td>Made in the current experiment</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Desalting column</td>
			<td>Thermo scientific</td>
			<td>WC321753</td>
			<td></td>
		</tr>
		<tr>
			<td>Deoxycholic acid</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>G818BA0029</td>
			<td></td>
		</tr>
		<tr>
			<td>DH5&#945; competent cells</td>
			<td>Made in the current experiment</td>
			<td></td>
			<td>E.coli DH5&#945;</td>
		</tr>
		<tr>
			<td>&#946;-D-maltoside</td>
			<td>Beijing Scolario Science and Tech Co.Ltd</td>
			<td>S818</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>E104BA0029</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycine</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>161BA0031</td>
			<td></td>
		</tr>
		<tr>
			<td>HEPES</td>
			<td>Beijing solaribo science and technology Co.Ltd</td>
			<td>H8090</td>
			<td></td>
		</tr>
		<tr>
			<td>LiCl</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>H209BA0003</td>
			<td></td>
		</tr>
		<tr>
			<td>MES</td>
			<td>Beijing solaribo science and technology Co.Ltd</td>
			<td>M8019</td>
			<td></td>
		</tr>
		<tr>
			<td>MiraCloth</td>
			<td>EMD Milipore Corp/MERCK kgAa Darmstadt, Germenay</td>
			<td>3429963</td>
			<td>Quick filtration material filter</td>
		</tr>
		<tr>
			<td>MgCl2</td>
			<td>Beijing solaribo science and technology Co.Ltd</td>
			<td>20200819</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>H324BA0003</td>
			<td></td>
		</tr>
		<tr>
			<td>NP40</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>N8030</td>
			<td></td>
		</tr>
		<tr>
			<td>Protein inhibitor cocktail</td>
			<td>Beijing Scolario Science and Tech Co.Ltd</td>
			<td>S3450</td>
			<td></td>
		</tr>
		<tr>
			<td>PVDF</td>
			<td>BIO-RAD</td>
			<td>5820172</td>
			<td></td>
		</tr>
		<tr>
			<td>SDS</td>
			<td>Beijing Scolario Science and Tech Co.Ltd</td>
			<td>S1015</td>
			<td></td>
		</tr>
		<tr>
			<td>Silwet</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>S9430</td>
			<td></td>
		</tr>
		<tr>
			<td>Streptavidin-C1-conjugated magnetic beads</td>
			<td>Enriching Biotechnology</td>
			<td>7E511E1</td>
			<td>Magnetic beads</td>
		</tr>
		<tr>
			<td>TEMED</td>
			<td>Servicebio</td>
			<td>G2056</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>GB03BA007</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-HCl</td>
			<td>Sangon Biotech (Shanghai) Co.Ltd</td>
			<td>F828BA0020</td>
			<td></td>
		</tr>
		<tr>
			<td>Tryptone</td>
			<td>Thermo scientific</td>
			<td>LP0042</td>
			<td></td>
		</tr>
	</tbody>
</table>

airid-based proximity labeling for protein-protein interaction in plants

In mammalian cells and plants, proximity labeling (PL) approaches using modified ascorbate peroxidase (APEX) or the Escherichia coli biotin ligase BirA (known as BioID) have proven successful in identifying protein-protein interactions (PPIs). APEX, BioID, and TurboID, a revised version of BioID have some restrictions in addition to being valuable technologies. The recently developed AirID, a novel version of BioID for proximity identification in protein-protein interactions, overcame these restrictions. Previously, AirID has been used in animal models, while the current study demonstrates the use of AirID in plants, and the results confirmed that AirID performs better in plant systems as compared to other PL enzymes such as BioID and TurboID for protein labeling that are proximal to the target proteins. Here is a step-by-step protocol for identifying protein interaction partners using AT4G18020 (APRR2) protein as a model. The methods describe the construction of vector, the transformation of construct through agroinfiltration, biotin transformation, extraction of proteins, and enrichment of biotin-labeled proteins through affinity purification technique. The results conclude that AirID is a novel and ideal enzyme for analyzing PPIs in plants. The method can be applied to study other proteins in plants.

According to previous research, the cucumber gene APRR2 is the candidate gene that controls white immature fruit color8. Here, a protocol was developed using AirID as a proximity labeling enzyme to find the interacting partner protein of APRR2 in cucumber. The construct was transferred to the cucumber leaves, and after 36 h post infiltration, biotin was transferred. After 48 h the samples were taken for western blot analysis to confirm the successful transformation. The proteins ...

Watch this Scientific Journal Video about AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants at JoVE.com

AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants

Here, we present a step-by-step protocol for performing the proximity labeling (PL) experiment in cucumber (Cucumis sativus L.) using AT4G18020 (APRR2)-AirID protein as a model. The method describes the construction of a vector, the transformation of a construct through agroinfiltration, biotin infiltration, protein extraction, and purification of biotin-labeled proteins through affinity purification technique.

In mammalian cells and plants, proximity labeling (PL) approaches using modified ascorbate peroxidase (APEX) or the Escherichia coli biotin ligase BirA ...

AirID는 단백질-단백질 상호 작용에 유용한 효소이며, 다른 근접 표지 효소보다 시간이 걸리는 공정에서 독성과 오류가 적습니다. APRR2-AirID 단백질을 모델로 사용하여 오이 또는 Cucumis sativus에서 근접 표지 실험AT4G18020 수행하기 위한 단계별 프로토콜이 제시됩니다. 제 연구실의 Ahmad Zada, Imran Khan, Yuting Cheng 및 Min Zhang이 절차를 시연할 것입니다.Cucumis sativus 또는 오이 씨앗을 물에 옮기고 섭씨 50도에서 20 분 동안 배양 한 다음 12-16 시간 동안 페트리 플레이트의 여과지에 씨앗을 놓습니다. 나중에, 씨앗을 흙이 들어있는 화분에 옮기고 섭씨 23도의 기후 챔버에서 16 시간 빛과 18 시간 어두운 사진 촬영 기간 동안 3-4 주 동안 자랍니다. 2.5 마이크로리터의 플라스미드 EarleyGate 100을 agrobacterium tumefaciens GV3101 균주의 유능한 세포로 옮깁니다.튜브를 얼음 위에서 30분 동안 배양한 후 액체 질소로 3분 동안 옮깁니다. 열 충격을 주려면 튜브를 섭씨 37도의 인큐베이터로 5분 동안 옮깁니다. 그런 다음 튜브를 얼음 위에 2분 동안 올려 놓습니다.1개, Luria Bertani LB 매체의 000 마이크로리터를 열에 충격을 가한 agrobacterium 세포에 추가하십시오. 세포를 섭씨 30도, 118RPM에서 1시간 동안 배양한 후 섭씨 4도에서 2분 동안 3000G로 원심분리합니다. 그런 다음 800 마이크로 리터 상층액을 버리고 나머지 용액을 혼합하십시오.카나마이신과 겐타마이신을 각각 밀리리터당 50마이크로그램, 리팜피신 밀리리터당 25마이크로그램을 보충한 LB 배지에 접시에 담습니다. 접시를 섭씨 30도에서 48시간 동안 배양합니다. 접시에서 박테리아 군집을 집어 올리십시오.적절한 항생제를 보충한 LB 액체 배지에 넣고 섭씨 30도, 218RPM에서 36-48시간 동안 액체 LB를 배양합니다. 3, 4 섭씨 온도에 000 G에 원심분리의 2 분 후에, 600 나노미터 파장에 것에 광학 조밀도를 조정하기 위하여 agroinfiltration 완충기에 있는 펠릿을 재현탁시키십시오. 1 밀리리터의 바늘 없는 주사기를 사용하여 1.5 밀리리터의 재현탁 농균 접종물을 복축표면의 표피 전체에 침투시킵니다.8 시간 동안 어두운 조건에서 16 시간 동안 초당 제곱 미터 당 75 마이크로 몰 빛으로 섭씨 23 도의 기후 챔버에서 식물을 유지하십시오 36 시간 후, 이미 여과 된 잎에 5 마이크로 그램의 비오틴 1 밀리리터를 침투시킵니다. 4-12시간 동안 비오틴이 침투한 후 잎을 자르고 액체 질소로 빠르게 옮겨 단백질 분해를 방지하십시오. 유봉과 절구를 사용하여 잎을 갈고 pH 7.4의 PBS BSA 2ml를 빠르게 첨가한 다음 천천히 분쇄합니다.샘플 혼합물을 그 위에 놓인 빠른 여과 재료 필터를 통해 15밀리리터 원뿔형 튜브로 옮기고 튜브를 얼음 위에 유지합니다. 샘플을 2밀리리터 튜브로 옮깁니다. 1% 단백질 억제제 칵테일을 넣고 원심분리하기 전에 튜브를 위아래로 7-8번 돌려 내용물을 섞습니다.완료되면 상층액을 새 2밀리리터 튜브에 옮기고 10% 베타-D-말토사이드를 추가합니다. 20, 4 섭씨 온도에 10 분 동안 000 G에 분리하기 전에 5 분 동안 얼음에 관을 두십시오. 탈염 컬럼의 평형을 맞추려면 컬럼의 한쪽 면을 표시하고 표시된 면이 바깥쪽을 향하도록 원심분리합니다.란의 정상과 밑바닥 덮개를 제거하고 4 섭씨 온도에 1, 000 G에 2 분 동안 다시 분리기를 분리하십시오. 컬럼의 수집 튜브에서 액체 용액을 버리고 이전에 시연된 대로 원심분리를 통해 5mL의 PBS 완충액으로 튜브를 최소 5회 세척합니다. 1.5밀리리터 튜브에 있는 50마이크로리터의 스트렙타비딘-C1 복합 마그네틱 비드에 PBS BSA 1밀리리터를 넣고 잘 섞습니다.튜브를 자석 스탠드에 3분 동안 올려 튜브의 한쪽 쪽으로 비드를 흡수시킵니다. 반대쪽의 상층액을 버리고 이 PBS BSA 세척을 세 번 반복합니다. 비오틴화된 단백질을 농축하려면 컬럼에 2mL의 샘플을 추가합니다.섭씨 4도에서 8분 동안 1000G에서 컬럼을 원심분리한 후 1mL의 탈염 단백질 추출물을 50마이크로리터의 Streptavidin-C1 복합 마그네틱 비드에 추가합니다. 자성 비드가 들어있는 튜브를 정상 속도와 실온에서 30분 동안 회전기에 올려 용액을 완전히 혼합합니다. 그런 다음 실온에서 3분 동안 또는 튜브의 한쪽에 모일 때까지 마그네틱 랙에 비드를 놓습니다.상층액을 부드럽게 제거하고 세척 완충액 1 1 밀리리터를 추가합니다. 로테이터에서 튜브를 2분 동안 회전시키고 마그네틱 랙에서 수행한 단계를 반복하여 상층액을 제거합니다. 마찬가지로, 이전에 설명한 대로 1밀리리터의 세척 버퍼 2와 3으로 세척을 수행합니다.다음으로, 1.7 밀리 몰 트리스 하이드로 클로라이드 50 밀리리터를 추가하고 마그네틱 랙에서 수행 된 단계를 반복하여 세척 버퍼에서 세제를 제거합니다. 50 밀리몰 암모늄 중탄산염 완충액 1 밀리리터를 사용하여 각각 2 분씩 6 회 세척하십시오. 다 익으면 염산삼중염산염 50밀리몰, 수크로스 12%, 라우릴리튬 황산염 2, 디티오트레이톨 1.5%가 함유된 단백질 추출물 50마이크로리터를 비드에 넣고 튜브를 섭씨 100도의 인큐베이터에 5분간 넣어 열충격을 줍니다.마그네틱 랙에서 단계를 반복한 후 LCMS/MS 분석을 위해 상층액을 섭씨 영하 80도에서 보관합니다. 추출된 단백질의 웨스턴 블롯 분석은 침윤된 오이 잎에서 성공적인 단백질 발현 및 비오틴화를 보여주었습니다. 대조군에 비해 표지된 단백질과 추가 밴드의 발현 수준이 높을수록 AirID에 의한 관심 유전자의 표적 단백질의 성공적인 태깅이 확인되었습니다.결과는 모든 형질전환된 샘플에서 Anti-Flag 항체가 있는 표적 밴드를 보여주는 Anti-Flag 및 Anti-Mass 항체를 사용하여 확인되었습니다. 스트렙타비딘-C1 접합 마그네틱 비드로 비오틴화된 단백질을 농축한 후 크기가 다른 여러 단백질을 관찰했습니다. 모든 결과에서 AirID는 식물의 단백질-단백질 상호 작용을 분석하기 위한 새롭고 이상적인 효소라는 결론을 내렸습니다.근접 표지 실험 중에는 5마이크로몰 비오틴과 HR 지속 시간이 이상적이라는 점을 기억하는 것이 중요합니다. 이 프로토콜은 잎 샘플 준비, 프리비오틴 제거, 추출물 단백질 정량화 및 비오틴화 단백질 농축을 포함하여 식물에서 AirID 기반 근접 라벨링의 단계별 설정을 간략하게 설명합니다. AirID는 PPI 독립적 비오틴화 정확도를 함께 향상시킬 것으로 예상됩니다.AirID는 식물의 PPI 분석에 이상적이라는 결론을 내렸습니다.

Research

JoVE Journal

Biology

Bimolecular Fluorescence Complementation (BiFC) Assay for Protein-Protein Interaction in Onion Cells Using the Helios Gene Gun

Bimolecular Fluorescence Complementation BiFC Assay for Protein-Protein Interaction in Onion Cells Using the Helios Gene Gun

헬리 우스 진 건을 사용하여 양파 세포에서 단백질 - 단백질 상호 작용에 대한 Bimolecular 형광 Complementation (BiFC) 어세이

Investigation of gene function in diverse organisms relies on knowledge of how the gene products interact with each other in their normal cellular ...

연구

생물학

Imaging Protein-protein Interactions in vivo

Imaging Protein-protein Interactions in vivo

이미징 단백질 - 단백질 상호 작용 생체내에</em

Protein-protein interactions are a hallmark of all essential cellular processes. However, many of these interactions are transient, or energetically weak, ...

A Liquid Phase Affinity Capture Assay Using Magnetic Beads to Study Protein-Protein Interaction: The Poliovirus-Nanobody Example

단백질 - 단백질 상호 작용 연구를 자성 비즈를 사용하여 액상 동질 캡처 분석 : Poliovirus-Nanobody의 예

In this article, a simple, quantitative, liquid phase affinity capture assay is presented. Provided that one protein can be tagged and another protein ...

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

을 바탕으로 단백질 - 단백질 상호 작용의 확인에 프로토콜<sup&gt; 15</sup&gt; N 신진 대사 라벨링, Immunoprecipitation, 수량 질량 분석법 및 선호도 변조

Protein-protein interactions are fundamental for many biological processes in the cell. Therefore, their characterization plays an important role in ...

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction

이분자 형광 보완의 유세포 분석 : 단백질 - 단백질 상호 작용을 연구하는 높은 처리량 정량 방법

Among methods to study protein-protein interaction inside cells, Bimolecular Fluorescence Complementation (BiFC) is relatively simple and sensitive. BiFC ...

Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification (TAP)

Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification TAP

단백질 - 단백질 상호 작용을 확인<em&gt; 초파리</em탠덤 선호도 정화로&gt; 성인 머리 (TAP)

Genetic screens conducted using Drosophila melanogaster (fruit fly) have made numerous milestone discoveries in the advance of biological sciences. ...

Investigating Protein-protein Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer

생물 발광 공명 에너지 전달을 이용하여 살아있는 세포에서 단백질 - 단백질 상호 작용을 조사

Assays based on Bioluminescence Resonance Energy Transfer (BRET) provide a sensitive and reliable means to monitor protein-protein interactions in live ...

Measuring Spatial and Temporal Ca2+ Signals in Arabidopsis Plants

Measuring Spatial and Temporal Ca2+ Signals in Arabidopsis Plants

측정 공간적 칼슘<sup&gt; 2 +</sup애기 장대 식물에서&gt; 신호

Developmental and environmental cues induce Ca2+ fluctuations in plant cells. Stimulus-specific spatial-temporal Ca2+ patterns are sensed by cellular Ca2+ ...

Identifying Protein-protein Interaction Sites Using Peptide Arrays

펩타이드 배열을 사용하여 단백질 - 단백질 상호 작용 사이트를 확인

Protein-protein interactions mediate most of the processes in the living cell and control homeostasis of the organism. Impaired protein interactions may ...

Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay (PCA) in Living Cells

Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay PCA in Living Cells

살아있는 세포의 단백질 조각 보완 분석 (PCA)에 의해 게놈 전체 단백질 - 단백질 상호 작용 상영

Proteins are the building blocks, effectors and signal mediators of cellular processes. A protein&#8217;s function, regulation and localization often ...