Protein Science最新文献_第7页

Distinct substrate specificities of the three catalytic subunits of the Trichomonas vaginalis proteasome. 阴道毛滴虫蛋白酶体三个催化亚基不同的底物特异性。

IF 4.5 3区生物学

Protein Science Pub Date : 2024-12-01 DOI: 10.1002/pro.5225

Pavla Fajtova, Brianna M Hurysz, Yukiko Miyamoto, Mateus Sá M Serafim, Zhenze Jiang, Julia M Vazquez, Diego F Trujillo, Lawrence J Liu, Urvashi Somani, Jehad Almaliti, Samuel A Myers, Conor R Caffrey, William H Gerwick, Dustin L McMinn, Christopher J Kirk, Evzen Boura, Lars Eckmann, Anthony J O'Donoghue

{"title":"Distinct substrate specificities of the three catalytic subunits of the Trichomonas vaginalis proteasome.","authors":"Pavla Fajtova, Brianna M Hurysz, Yukiko Miyamoto, Mateus Sá M Serafim, Zhenze Jiang, Julia M Vazquez, Diego F Trujillo, Lawrence J Liu, Urvashi Somani, Jehad Almaliti, Samuel A Myers, Conor R Caffrey, William H Gerwick, Dustin L McMinn, Christopher J Kirk, Evzen Boura, Lars Eckmann, Anthony J O'Donoghue","doi":"10.1002/pro.5225","DOIUrl":"10.1002/pro.5225","url":null,"abstract":"The protozoan parasite Trichomonas vaginalis (Tv) causes trichomoniasis, the most common non-viral sexually transmitted infection in the world. Although Tv has been linked to significant health complications, only two closely related 5-nitroimidazole drugs are approved for its treatment. The emergence of resistance to these drugs and lack of alternative treatment options poses an increasing threat to public health, making development of novel anti-Trichomonas compounds an urgent need. The proteasome, a critical enzyme complex found in all eukaryotes has three catalytic subunits, β1, β2, and β5 and has been validated as a drug target to treat trichomoniasis. With the goal of developing tools to study the Tv proteasome, we isolated the enzyme complex and identified inhibitors that preferentially inactivate either one or two of the three catalytic subunits. Using a mass spectrometry-based peptide digestion assay, these inhibitors were used to define the substrate preferences of the β1, β2 and β5 subunits. Subsequently, three model fluorogenic substrates were designed, each specific for one of the catalytic subunits. This novel substrate profiling methodology will allow for individual subunit characterization of other proteasomes of interest. Using the new substrates, we screened a library of 284 peptide epoxyketone inhibitors against Tv and determined the subunits targeted by the most active compounds. The data show that inhibition of the Tv β5 subunit alone is toxic to the parasite. Taken together, the optimized proteasome subunit substrates will be instrumental for understanding the molecular determinants of proteasome specificity and for accelerating drug development against trichomoniasis.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"33 12","pages":"e5225"},"PeriodicalIF":4.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11590128/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142716977","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Comparative studies of seafood and reptile α- and β-parvalbumins. 海产品和爬行动物 α- 和 β-副缬氨酸的比较研究。

IF 4.5 3区生物学

Protein Science Pub Date : 2024-12-01 DOI: 10.1002/pro.5226

Andrea O'Malley, Joshua M Ray, Patrycja Kitlas, Thimo Ruethers, A Brenda Kapingidza, Tomasz Cierpicki, Andreas Lopata, Krzysztof Kowal, Maksymilian Chruszcz

{"title":"Comparative studies of seafood and reptile α- and β-parvalbumins.","authors":"Andrea O'Malley, Joshua M Ray, Patrycja Kitlas, Thimo Ruethers, A Brenda Kapingidza, Tomasz Cierpicki, Andreas Lopata, Krzysztof Kowal, Maksymilian Chruszcz","doi":"10.1002/pro.5226","DOIUrl":"10.1002/pro.5226","url":null,"abstract":"Small calcium-binding proteins such as parvalbumins (PVs) are major seafood and fish allergens. However, the impact of structural changes on their capacity to bind IgE has not been studied in detail. Therefore, fish and reptilian PVs, as well as human α-PV, were selected for biochemical, structural, and IgE binding studies. Likely due to their high solubility, crystallization proved difficult, so additional techniques were used to promote crystallization of the proteins. Novel crystal structures were determined for human PV, cod allergen Gad m 1.0201, saltwater crocodile allergen Cro p 1.0101, and the α-PV from thornback ray. β-PVs are considered the major fish allergens, while α-PVs are rarely categorized as allergens. To explain these differences, the results of structural and IgE binding studies were combined. This approach allowed us to provide new insight into IgE binding epitopes present on PVs, focusing on cross-reactivity among the selected α- and β-PVs. In addition, we have shown that these proteins display remarkable thermal stability across a range of pH conditions, which is relevant in the case of food allergens and food processing. Moreover, it is shown that the presence of calcium cations is critical for stability of the studied PVs via their protein folding, which has an impact on the formation of IgE binding epitopes. These studies shows the stability of fish and reptile PV allergens, and it allows for further evaluation of their IgE cross-reactivity.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"33 12","pages":"e5226"},"PeriodicalIF":4.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11586863/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710932","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Special issue title: The Protein Society 38th Annual Symposium, July 2024, Vancouver, Canada. 特刊标题：蛋白质学会第38届年会，2024年7月，加拿大温哥华。

IF 4.5 3区生物学

Protein Science Pub Date : 2024-12-01 DOI: 10.1002/pro.5207

引用次数: 0

Impact of primary sequence changes on the self-association properties of mammalian cystathionine beta-synthase enzymes. 主序列变化对哺乳动物胱硫醚 beta 合成酶自结合特性的影响。

IF 4.5 3区生物学

Protein Science Pub Date : 2024-12-01 DOI: 10.1002/pro.5223

Hyung-Ok Lee, Kushol Gupta, Liqun Wang, Roland L Dunbrack, Tomas Majtan, Warren D Kruger

{"title":"Impact of primary sequence changes on the self-association properties of mammalian cystathionine beta-synthase enzymes.","authors":"Hyung-Ok Lee, Kushol Gupta, Liqun Wang, Roland L Dunbrack, Tomas Majtan, Warren D Kruger","doi":"10.1002/pro.5223","DOIUrl":"10.1002/pro.5223","url":null,"abstract":"Cystathionine beta-synthase (CBS) is an evolutionarily conserved enzyme that plays a key role in mammalian sulfur amino acid biochemistry, mutations in which are the cause of classical homocystinuria (HCU), an inborn error of metabolism. Although there is agreement in the literature that CBS is a homomultimer, its precise structure is a source of confusion. Here, we performed a series of experiments examining the quaternary structure of various wild-type and mutant CBS enzymes using a combination of native gel electrophoresis, in situ activity assays, analytical ultracentrifugation, and gel filtration. Our data show that recombinantly expressed and purified full-length wild-type human CBS enzyme (hCBS) and HCU-causing variants (p.P422L, p.I435T, and p.R125Q CBS) form high molecular weight assemblies that are consistent with the properties expected of a filament. The filament is enzymatically active, and its size is sensitive to protein concentration. This behavior contrasts sharply with hCBS enzymes containing small deletions within the Bateman domain, which form stable tetramers and octamers regardless of concentration. Examination of liver lysates from humans and mice confirms the existence of enzymatically active high molecular weight aggregates in vivo, but also shows that these aggregates are specific to human CBS and do not occur in mice. Molecular modeling using AlphaFold2 suggests that these experimentally observed differences may be explained by subtle differences in the interaction mediated by the Bateman domains. Our results show that small differences in amino acid sequence can cause large differences in the size and shape of CBS multimers.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"33 12","pages":"e5223"},"PeriodicalIF":4.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11568414/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142644525","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The ubiquitous pyridoxal 5'-phosphate-binding protein is also an RNA-binding protein. 无处不在的 5'-磷酸吡哆醛结合蛋白也是一种 RNA 结合蛋白。

IF 4.5 3区生物学

Protein Science Pub Date : 2024-12-01 DOI: 10.1002/pro.5242

Claudio Graziani, Anna Barile, Alessia Parroni, Martino Luigi di Salvo, Irene De Cecio, Teresa Colombo, Jill Babor, Valérie de Crécy-Lagard, Roberto Contestabile, Angela Tramonti

{"title":"The ubiquitous pyridoxal 5'-phosphate-binding protein is also an RNA-binding protein.","authors":"Claudio Graziani, Anna Barile, Alessia Parroni, Martino Luigi di Salvo, Irene De Cecio, Teresa Colombo, Jill Babor, Valérie de Crécy-Lagard, Roberto Contestabile, Angela Tramonti","doi":"10.1002/pro.5242","DOIUrl":"10.1002/pro.5242","url":null,"abstract":"The pyridoxal 5'-phosphate binding protein (PLP-BP) is believed to play a crucial role in PLP homeostasis, which may explain why it is found in living organisms from all kingdoms. Escherichia coli YggS is the most studied homolog, but human PLP-BP has also attracted much attention because variants of this protein are responsible for a severe form of B6-responsive neonatal epilepsy. Yet, how PLP-BP is involved in PLP homeostasis, and thus what its actual function is in cellular metabolism, is entirely unknown. The present study shows that YggS binds RNA and that the strength of this interaction is modulated by PLP. A key role in RNA binding is clearly played by Lys137, an invariant residue located on a protein loop away from the PLP binding site, whose importance has been highlighted previously. The interaction with RNA is evidently conserved, since it is also observed with human PLP-BP. The RNA binding site, which is apparently located at the entrance of the PLP-binding site, is also evolutionarily conserved. It is therefore reasonable to assume that PLP, by defining the conformation of the protein, determines the RNA binding affinity. RNA-seq analysis of RNA co-purified with or captured by YggS revealed SsrA and RnpB RNAs, respectively involved in trans-translation and tRNA maturation, as the major molecular components. This work opens up new horizons for the function of the PLP-BP, which could be related to its interaction with RNA and modulated by PLP, and thus play a role in an as yet unknown regulatory mechanism.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"33 12","pages":"e5242"},"PeriodicalIF":4.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11602438/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142740256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The transmission of mutation effects in a multiprotein machine: A comprehensive metadynamics study of the cardiac thin filament. 多蛋白机器中突变效应的传递：心脏细丝的综合元动力学研究

IF 4.5 3区生物学

Protein Science Pub Date : 2024-12-01 DOI: 10.1002/pro.5215

Krishna Prasad Ghanta, Romi L Castillo, Jil C Tardiff, Steven D Schwartz

{"title":"The transmission of mutation effects in a multiprotein machine: A comprehensive metadynamics study of the cardiac thin filament.","authors":"Krishna Prasad Ghanta, Romi L Castillo, Jil C Tardiff, Steven D Schwartz","doi":"10.1002/pro.5215","DOIUrl":"10.1002/pro.5215","url":null,"abstract":"The binding of Ca2+ ions within the troponin core of the cardiac thin filament (CTF) regulates normal contraction and relaxation. Mutations within the troponin complexes are known to alter normal functions and result in the eventual development of cardiomyopathy. However, despite the importance of the problem, detailed microscopic knowledge of the mechanism of pathogenic effect of point mutations and their effects on the conformational free energy surface of CTF remains elusive. Mutations are known to transmit their effects hundreds of angstroms along this protein complex and between different component proteins. To explore the impact of point mutations on the conformational free energy barrier between the closed and blocked state of CTF, and to understand the transmission of mutation, we have carried out metadynamics simulations for the wild-type (WT) and two mutants (cardiac troponin T Arg92Trp (R92W) and Arg92Leu (R92L)). Specifically, we have investigated the conformational modification of the tropomyosin (Tm) and the troponin (Tn) complex during the closed-to-blocked state transition for both the WT and two hypertrophic cardiomyopathy causing mutations. Our calculations demonstrated that mutations within the cardiac troponin T (cTnT) protein alter conformational properties of the Tm and the other proteins of the Tn complex as well as the Ca2+ binding affinity of the cTnC protein through the indirect mediation of cardiac troponin I (cTnI). Importantly, the data revealed a significant influence of the mutations on the conformational transition free energy barriers for both the Tm and cTnC proteins. Furthermore, we found both mutations independently alter the free energy barrier of transitions of cTnT. Such alteration in the free energy upon mutation of one protein in a complex, allosterically affects the others through structural and dynamical changes, leading to a pathogenic effect on the function of the thin filament.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"33 12","pages":"e5215"},"PeriodicalIF":4.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11568392/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142644542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Preferential binding of ADP-bound mitochondrial HSP70 to the nucleotide exchange factor GRPEL1 over GRPEL2. ADP 结合的线粒体 HSP70 与核苷酸交换因子 GRPEL1 的结合优于 GRPEL2。

IF 4.5 3区生物学

Protein Science Pub Date : 2024-11-01 DOI: 10.1002/pro.5190

Pooja Manjunath, Gorazd Stojkovič, Liliya Euro, Svetlana Konovalova, Sjoerd Wanrooij, Kristian Koski, Henna Tyynismaa

{"title":"Preferential binding of ADP-bound mitochondrial HSP70 to the nucleotide exchange factor GRPEL1 over GRPEL2.","authors":"Pooja Manjunath, Gorazd Stojkovič, Liliya Euro, Svetlana Konovalova, Sjoerd Wanrooij, Kristian Koski, Henna Tyynismaa","doi":"10.1002/pro.5190","DOIUrl":"10.1002/pro.5190","url":null,"abstract":"Human nucleotide exchange factors GRPEL1 and GRPEL2 play pivotal roles in the ADP-ATP exchange within the protein folding cycle of mitochondrial HSP70 (mtHSP70), a crucial chaperone facilitating protein import into the mitochondrial matrix. Studies in human cells and mice have indicated that while GRPEL1 serves as an essential co-chaperone for mtHSP70, GRPEL2 has a role regulated by stress. However, the precise structural and biochemical mechanisms underlying the distinct functions of the GRPEL proteins have remained elusive. In our study, we present evidence revealing that ADP-bound mtHSP70 exhibits remarkably higher affinity for GRPEL1 compared to GRPEL2, with the latter experiencing a notable decrease in affinity upon ADP binding. Additionally, Pi assay showed that GRPEL1, but not GRPEL2, enhanced the ATPase activity of mtHSP70. Utilizing Alphafold modeling, we propose that the interaction between GRPEL1 and mtHSP70 can induce the opening of the nucleotide binding cleft of the chaperone, thereby facilitating the release of ADP, whereas GRPEL2 lacks this capability. Additionally, our findings suggest that the redox-regulated Cys87 residue in GRPEL2 does not play a role in dimerization but rather reduces its affinity for mtHSP70. Our findings on the structural and functional disparities between GRPEL1 and GRPEL2 may have implications for mitochondrial protein folding and import processes under varying cellular conditions.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"33 11","pages":"e5190"},"PeriodicalIF":4.5,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11500471/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142506691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Unraveling the molecular determinants of a rare human mitochondrial disorder caused by the P144L mutation of FDX2. 揭示由 FDX2 的 P144L 突变引起的罕见人类线粒体疾病的分子决定因素。

IF 4.5 3区生物学

Protein Science Pub Date : 2024-11-01 DOI: 10.1002/pro.5197

Deborah Grifagni, Davide Doni, Bianca Susini, Bruno M Fonseca, Ricardo O Louro, Paola Costantini, Simone Ciofi-Baffoni

{"title":"Unraveling the molecular determinants of a rare human mitochondrial disorder caused by the P144L mutation of FDX2.","authors":"Deborah Grifagni, Davide Doni, Bianca Susini, Bruno M Fonseca, Ricardo O Louro, Paola Costantini, Simone Ciofi-Baffoni","doi":"10.1002/pro.5197","DOIUrl":"10.1002/pro.5197","url":null,"abstract":"Episodic mitochondrial myopathy with or without optic atrophy and reversible leukoencephalopathy (MEOAL) is a rare, orphan autosomal recessive disorder caused by mutations in ferredoxin-2 (FDX2), which is a [2Fe-2S] cluster-binding protein participating in the formation of iron-sulfur clusters in mitochondria. In this biosynthetic pathway, FDX2 works as electron donor to promote the assembly of both [2Fe-2S] and [4Fe-4S] clusters. A recently identified missense mutation of MEOAL is the homozygous mutation c.431C>T (p.P144L) described in six patients from two unrelated families. This mutation alters a highly conserved proline residue located in a loop of FDX2 that is distant from the [2Fe-2S] cluster. How this Pro to Leu substitution damages iron-sulfur cluster biosynthesis is unknown. In this work, we have first compared the structural, dynamic, cluster binding and redox properties of WT and P144L [2Fe-2S] FDX2 to have clues on how the pathogenic P144L mutation can perturb the FDX2 function. Then, we have investigated the interaction of both WT and P144L [2Fe-2S] FDX2 with its physiological electron donor, ferredoxin reductase FDXR, comparing their electron transfer efficiency and protein-protein recognition patterns. Overall, the data indicate that the pathogenic P144L mutation negatively affects the FDXR-dependent electron transfer pathway from NADPH to FDX2, thereby reducing the capacity of FDX2 in assembling both [2Fe-2S] and [4Fe-4S] clusters. Our study also provided solid molecular evidences on the functional role of the C-terminal tail of FDX2 in the electron transfer between FDX2 and FDXR.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"33 11","pages":"e5197"},"PeriodicalIF":4.5,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11515921/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142522795","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Monomer unfolding of a bacterial ESCRT-III superfamily member is coupled to oligomer disassembly. 细菌 ESCRT-III 超家族成员的单体折叠与寡聚体解体相关联。

IF 4.5 3区生物学

Protein Science Pub Date : 2024-11-01 DOI: 10.1002/pro.5187

Ndjali Quarta, Tika Ram Bhandari, Martin Girard, Nadja Hellmann, Dirk Schneider

{"title":"Monomer unfolding of a bacterial ESCRT-III superfamily member is coupled to oligomer disassembly.","authors":"Ndjali Quarta, Tika Ram Bhandari, Martin Girard, Nadja Hellmann, Dirk Schneider","doi":"10.1002/pro.5187","DOIUrl":"10.1002/pro.5187","url":null,"abstract":"The inner membrane associated protein of 30 kDa (IM30), a member of the endosomal sorting complex required for transport (ESCRT-III) superfamily, is crucially involved in the biogenesis and maintenance of thylakoid membranes in cyanobacteria and chloroplasts. In solution, IM30 assembles into various large oligomeric barrel- or tube-like structures, whereas upon membrane binding it forms large, flat carpet structures. Dynamic localization of the protein in solution, to membranes and changes of the oligomeric states are crucial for its in vivo function. ESCRT-III proteins are known to form oligomeric structures that are dynamically assembled from monomeric/smaller oligomeric proteins, and thus these smaller building blocks must be assembled sequentially in a highly orchestrated manner, a still poorly understood process. The impact of IM30 oligomerization on function remains difficult to study due to its high intrinsic tendency to homo-oligomerize. Here, we used molecular dynamics simulations to investigate the stability of individual helices in IM30 and identified unstable regions that may provide structural flexibility. Urea-mediated disassembly of the IM30 barrel structures was spectroscopically monitored, as well as changes in the protein's tertiary and secondary structure. The experimental data were finally compared to a three-state model that describes oligomer disassembly and monomer unfolding. In this study, we identified a highly stable conserved structural core of ESCRT-III proteins and discuss the advantages of having flexible intermediate structures and their putative relevance for ESCRT-III proteins.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"33 11","pages":"e5187"},"PeriodicalIF":4.5,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11520248/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142522791","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Self-association and multimer formation in AtLEA4-5, a desiccation-induced intrinsically disordered protein from plants. AtLEA4-5（一种来自植物的干燥诱导型本征无序蛋白）的自结合和多聚体形成。

IF 4.5 3区生物学

Protein Science Pub Date : 2024-11-01 DOI: 10.1002/pro.5192

Paulette Sofía Romero-Pérez, Laura V Martínez-Castro, Alejandro Linares, Inti Arroyo-Mosso, Nuria Sánchez-Puig, Cesar L Cuevas-Velazquez, Shahar Sukenik, Adán Guerrero, Alejandra A Covarrubias

{"title":"Self-association and multimer formation in AtLEA4-5, a desiccation-induced intrinsically disordered protein from plants.","authors":"Paulette Sofía Romero-Pérez, Laura V Martínez-Castro, Alejandro Linares, Inti Arroyo-Mosso, Nuria Sánchez-Puig, Cesar L Cuevas-Velazquez, Shahar Sukenik, Adán Guerrero, Alejandra A Covarrubias","doi":"10.1002/pro.5192","DOIUrl":"10.1002/pro.5192","url":null,"abstract":"During seed maturation, plants may experience severe desiccation, leading to the accumulation of late embryogenesis abundant (LEA) proteins. These intrinsically disordered proteins also accumulate in plant tissues under water deficit. Functional roles of LEA proteins have been proposed based on in vitro studies, where monomers are considered as the functional units. However, the potential formation of homo-oligomers has been little explored. In this work, we investigated the potential self-association of Arabidopsis thaliana group 4 LEA proteins (AtLEA4) using in vitro and in vivo approaches. LEA4 proteins represent a compelling case of study due to their high conservation throughout the plant kingdom. This protein family is characterized by a conserved N-terminal region, with a high alpha-helix propensity and invitro protective activity, as compared to the highly disordered and low-conserved C-terminal region. Our findings revealed that full-length AtLEA4 proteins oligomerize and that both terminal regions are sufficient for self-association in vitro. However, the ability of both amino and carboxy regions of AtLEA4-5 to self-associate invivo is significantly lower than that of the entire protein. Using high-resolution and quantitative fluorescence microscopy, we were able to disclose the unreported ability of LEA proteins to form high-order oligomers in planta. Additionally, we found that high-order complexes require the simultaneous engagement of both terminal regions, indicating that the entire protein is needed to attain such structural organization. This research provides valuable insights into the self-association of LEA proteins in plants and emphasizes the role of protein oligomer formation.","PeriodicalId":20761,"journal":{"name":"Protein Science","volume":"33 11","pages":"e5192"},"PeriodicalIF":4.5,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11516066/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142522792","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0