Protein Engineering Design & Selection最新文献_第7页

Yeast surface display-based identification of ACE2 mutations that modulate SARS-CoV-2 spike binding across multiple mammalian species. 基于酵母表面展示的 ACE2 突变鉴定可调节多种哺乳动物的 SARS-CoV-2 穗状结合。

IF 2.6 4区生物学

Protein Engineering Design & Selection Pub Date : 2022-02-17 DOI: 10.1093/protein/gzab035

Pete Heinzelman, Jonathan C Greenhalgh, Philip A Romero

引用次数: 0

Structure-guided protein engineering of human cathepsin L for efficient collagenolytic activity. 结构导向的人组织蛋白酶L高效胶原溶解活性蛋白工程。

IF 2.4 4区生物学

Protein Engineering Design & Selection Pub Date : 2021-02-15 DOI: 10.1093/protein/gzab005

Debi Choudhury, Sampa Biswas

{"title":"Structure-guided protein engineering of human cathepsin L for efficient collagenolytic activity.","authors":"Debi Choudhury, Sampa Biswas","doi":"10.1093/protein/gzab005","DOIUrl":"https://doi.org/10.1093/protein/gzab005","url":null,"abstract":"Engineering precise substrate specificity of proteases advances the potential to use them in biotechnological and therapeutic applications. Collagen degradation, a physiological process mediated by collagenases, is an integral part of extracellular matrix remodeling and when uncontrolled, implicated in different pathological conditions. Lysosomal cathepsin-K cleaves triple helical collagen fiber, whereas cathepsin-L cannot do so. In this study, we have imparted collagenolytic property to cathepsin-L, by systematically engineering proline-specificity and glycosaminoglycans (GAG)-binding surface in the protease. The proline-specific mutant shows high specificity for prolyl-peptidic substrate but is incapable of cleaving collagen. Engineering a GAG-binding surface on the proline-specific mutant enabled it to degrade type-I collagen in the presence of chondroitin-4-sulfate (C4-S). We also present the crystal structures of proline-specific (1.4 Å) and collagen-specific (1.8 Å) mutants. Finally docking studies with prolyl-peptidic substrate (Ala-Gly-Pro-Arg-Ala) at the active site and a C4-S molecule at the GAG-binding site enable us to identify key structural features responsible for collagenolytic activity of cysteine cathepsins.","PeriodicalId":54543,"journal":{"name":"Protein Engineering Design & Selection","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2021-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/protein/gzab005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25566737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Hypomorphic mutations in human DNA ligase IV lead to compromised DNA binding efficiency, hydrophobicity and thermal stability. 人类DNA连接酶IV的半胚突变导致DNA结合效率、疏水性和热稳定性受损。

IF 2.4 4区生物学

Protein Engineering Design & Selection Pub Date : 2021-02-15 DOI: 10.1093/protein/gzab001

Eswar Reddy Maddi, Sathees C Raghavan, Ramanathan Natesh

{"title":"Hypomorphic mutations in human DNA ligase IV lead to compromised DNA binding efficiency, hydrophobicity and thermal stability.","authors":"Eswar Reddy Maddi, Sathees C Raghavan, Ramanathan Natesh","doi":"10.1093/protein/gzab001","DOIUrl":"https://doi.org/10.1093/protein/gzab001","url":null,"abstract":"Studies have shown that Lig4 syndrome mutations in DNA ligase IV (LigIV) are compromised in its function with residual level of double strand break ligation activity in vivo. It was speculated that Lig4 syndrome mutations adversely affect protein folding and stability. Though there are crystal structures of LigIV, there are no reports of crystal structures of Lig4 syndrome mutants and their biophysical characterization to date. Here, we have examined the conformational states, thermal stability, hydrophobicity and DNA binding efficiency of human DNA LigIV wild type and its hypomorphic mutants by far-UV circular dichroism, tyrosine and tryptophan fluorescence, and 1-anilino-8-naphthalene-sulfonate binding, dynamic light scattering, size exclusion chromatography, multi-angle light scattering and electrophoretic mobility shift assay. We show here that LigIV hypomorphic mutants have reduced DNA-binding efficiency, a shift in secondary structure content from the helical to random coil, marginal reduction in their thermal stability and increased hydrophobicity as compared to the wild-type LigIV.","PeriodicalId":54543,"journal":{"name":"Protein Engineering Design & Selection","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2021-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/protein/gzab001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25373436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Design and engineering of artificial metalloproteins: from de novo metal coordination to catalysis. 人造金属蛋白的设计与工程:从从头金属配位到催化。

IF 2.4 4区生物学

Protein Engineering Design & Selection Pub Date : 2021-02-15 DOI: 10.1093/protein/gzab003

Andreas S Klein, Cathleen Zeymer

引用次数: 9

Improved thermostability of proteinase K and recognizing the synergistic effect of Rosetta and FoldX approaches. 改善蛋白酶K的热稳定性，并认识到Rosetta和FoldX方法的协同效应。

IF 2.4 4区生物学

Protein Engineering Design & Selection Pub Date : 2021-02-15 DOI: 10.1093/protein/gzab024

Yang Zhao, Daixi Li, Xue Bai, Manjie Luo, Yan Feng, Yilei Zhao, Fuqiang Ma, Guang-Yu Yang

{"title":"Improved thermostability of proteinase K and recognizing the synergistic effect of Rosetta and FoldX approaches.","authors":"Yang Zhao, Daixi Li, Xue Bai, Manjie Luo, Yan Feng, Yilei Zhao, Fuqiang Ma, Guang-Yu Yang","doi":"10.1093/protein/gzab024","DOIUrl":"https://doi.org/10.1093/protein/gzab024","url":null,"abstract":"Proteinase K (PRK) is a proteolytic enzyme that has been widely used in industrial applications. However, poor stability has severely limited the uses of PRK. In this work, we used two structure-guided rational design methods, Rosetta and FoldX, to modify PRK thermostability. Fifty-two single amino acid conversion mutants were constructed based on software predictions of residues that could affect protein stability. Experimental characterization revealed that 46% (21 mutants) exhibited enhanced thermostability. The top four variants, D260V, T4Y, S216Q, and S219Q, showed improved half-lives at 69°C by 12.4-, 2.6-, 2.3-, and 2.2-fold that of the parent enzyme, respectively. We also found that selecting mutations predicted by both methods could increase the predictive accuracy over that of either method alone, with 73% of the shared predicted mutations resulting in higher thermostability. In addition to providing promising new variants of PRK in industrial applications, our findings also show that combining these programs may synergistically improve their predictive accuracy.","PeriodicalId":54543,"journal":{"name":"Protein Engineering Design & Selection","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2021-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39535240","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

Breakthroughs in computational design methods open up new frontiers for de novo protein engineering. 计算设计方法的突破为从头开始的蛋白质工程开辟了新的领域。

IF 2.4 4区生物学

Protein Engineering Design & Selection Pub Date : 2021-02-15 DOI: 10.1093/protein/gzab007

Ben A Meinen, Christopher D Bahl

引用次数: 9

Linking thermodynamics and measurements of protein stability. 把热力学和蛋白质稳定性的测量联系起来。

IF 2.4 4区生物学

Protein Engineering Design & Selection Pub Date : 2021-02-15 DOI: 10.1093/protein/gzab002

Kresten Lindorff-Larsen, Kaare Teilum

引用次数: 9

Engineering stable carbonic anhydrases for CO2 capture: a critical review. 工程稳定的碳酸酐酶用于二氧化碳捕获:一个重要的回顾。

IF 2.4 4区生物学

Protein Engineering Design & Selection Pub Date : 2021-02-15 DOI: 10.1093/protein/gzab021

Mirfath Sultana Mesbahuddin, Aravindhan Ganesan, Subha Kalyaanamoorthy

引用次数: 7

A versatile assay platform for enzymatic poly(ethylene-terephthalate) degradation. 一个通用的酶促聚对苯二甲酸乙酯降解测定平台。

IF 2.4 4区生物学

Protein Engineering Design & Selection Pub Date : 2021-02-15 DOI: 10.1093/protein/gzab022

Sebastian Weigert, Andreas Gagsteiger, Teresa Menzel, Birte Höcker

引用次数: 3

Rational engineering of an erythropoietin fusion protein to treat hypoxia. 合理设计促红细胞生成素融合蛋白治疗缺氧。

IF 2.4 4区生物学

Protein Engineering Design & Selection Pub Date : 2021-02-15 DOI: 10.1093/protein/gzab025

Jungmin Lee, Andyna Vernet, Nathalie G Gruber, Kasia M Kready, Devin R Burrill, Jeffrey C Way, Pamela A Silver

{"title":"Rational engineering of an erythropoietin fusion protein to treat hypoxia.","authors":"Jungmin Lee, Andyna Vernet, Nathalie G Gruber, Kasia M Kready, Devin R Burrill, Jeffrey C Way, Pamela A Silver","doi":"10.1093/protein/gzab025","DOIUrl":"https://doi.org/10.1093/protein/gzab025","url":null,"abstract":"Erythropoietin enhances oxygen delivery and reduces hypoxia-induced cell death, but its pro-thrombotic activity is problematic for use of erythropoietin in treating hypoxia. We constructed a fusion protein that stimulates red blood cell production and neuroprotection without triggering platelet production, a marker for thrombosis. The protein consists of an anti-glycophorin A nanobody and an erythropoietin mutant (L108A). The mutation reduces activation of erythropoietin receptor homodimers that induce erythropoiesis and thrombosis, but maintains the tissue-protective signaling. The binding of the nanobody element to glycophorin A rescues homodimeric erythropoietin receptor activation on red blood cell precursors. In a cell proliferation assay, the fusion protein is active at 10-14 M, allowing an estimate of the number of receptor-ligand complexes needed for signaling. This fusion protein stimulates erythroid cell proliferation in vitro and in mice, and shows neuroprotective activity in vitro. Our erythropoietin fusion protein presents a novel molecule for treating hypoxia.","PeriodicalId":54543,"journal":{"name":"Protein Engineering Design & Selection","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2021-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39835737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1