Preservation of growth factor sensitivity and bioactivity(e.g.,bone morphogenetic protein-2(BMP-2))post-immobilization to tissue engineering scaffolds remains a great challenge.Here,we develop a stable and soft surfac...Preservation of growth factor sensitivity and bioactivity(e.g.,bone morphogenetic protein-2(BMP-2))post-immobilization to tissue engineering scaffolds remains a great challenge.Here,we develop a stable and soft surface modification strategy to address this issue.BMP-2(a model growth factor)is covalently immobilized onto homogeneous poly(glycidyl methacrylate)(PGMA)polymer brushes which are grafted onto substrate surfaces(Au,quartz glass,silica wafer,or common biomaterials)via surface-initiated atom transfer radical polymerization.This surface modification method multiplies the functionalized interfacial area;it is simple,fast,gentle,and has little effect on the loaded protein owing to the cilia motility.The immobilized BMP-2(i-BMP-2)on the surface of homogeneous PGMA polymer brushes exhibits excellent bioactivity(-87%bioactivity of free BMP-2 in vitro and 20%-50%higher than scaffolds with free BMP-2 in vivo),with conformation and secondary structure well-preserved after covalent immobilization and ethanol sterilization.Moreover,the osteogenic activity of i-BMP-2 on the nanoline pattern(PGMA-poly(N-isopropylacrylamide))shows-110%bioactivity of free BMP-2.This is superior compared to conventional protein covalent immobilization strategies in terms of both bioactivity preservation and therapeutic efficacy.PGMA polymer brushes can be used to modify surfaces of different tissue-engineered scaffolds,which facilitates in situ immobilization of growth factors,and accelerates repair of a wide range of tissue types.展开更多
Recently,multifunctional polymers for protein conjugation have been facilely synthesized through the multicomponent reactions(MCRs).In this mini-review,current progress in the generation of multifuncti onal polymers w...Recently,multifunctional polymers for protein conjugation have been facilely synthesized through the multicomponent reactions(MCRs).In this mini-review,current progress in the generation of multifuncti onal polymers with protei n-reactive groups via MCRs is summarized.These mult functional polymers react with model therapeutic proteins,forming multifunctional polymer-protein conjugates,which are prototypes of sophisticated theranostic agents and antibacterial vaccine candidates.In comparison with the traditional multi-step synthesis,the preparation of multifunctional polymers for protein conjugation through MCRs is straightforward and convenient.Due to these properties,MCRs have the potential to become a new general strategy to achieve polymer-protein conjugates for biological and medical applications.展开更多
Over the past 20 years,great efforts have been invested in developing site-specific approaches to protein modification to dissect protein functions directly and accurately.Here,we report a proximitytriggered group tra...Over the past 20 years,great efforts have been invested in developing site-specific approaches to protein modification to dissect protein functions directly and accurately.Here,we report a proximitytriggered group transfer strategy from a sulfonium warhead to a Cysteine(Cys)residue of the target protein.With a guiding ligand,cargoes could be transferred selectively from a sulfonium center onto the Cys residue in the vicinity of their binding interface.The successful thalidomide transfer of sulfonium 1-X could be applied intracellularly for epidermal growth factor receptor degradation,highlighting the potential of group transfer strategy as a suite of chemical biology studies,including cell imaging,protein profiling,and protein degradation by simply employing different transferrable groups.展开更多
Artificial synthesis and site-specific modification of peptides and proteins has evolved into an indispensable tool for protein engineers and chemical biologists. Chemical and enzymatic approaches to peptide ligation ...Artificial synthesis and site-specific modification of peptides and proteins has evolved into an indispensable tool for protein engineers and chemical biologists. Chemical and enzymatic approaches to peptide ligation are important alternatives of recombinant DNA technology for protein synthesis and modification. Although as old as that of chemical procedures, enzyme-mediated peptide ligation is far less developed than that of chemical counterpart due to the difficult availability of peptide ligase.Fortunately, this situation has been changed slowly with the fast development of biological techniques. In the past decades, several natural peptide ligases have been discovered. Protein engineering to improve the ligation efficiencies of the natural peptide ligase and to reverse the functionality of protease provide more powerful peptide ligases. In this review, the advances of enzyme-mediated peptide ligation and their application in protein synthesis and modification will be discussed.展开更多
Making peptide bonds is tightly controlled by genetic code and machinery which includes cofactors,ATP,and RNAs.In this regard,the stand-alone and genetic-code-independent peptide ligases constitute a new family of ren...Making peptide bonds is tightly controlled by genetic code and machinery which includes cofactors,ATP,and RNAs.In this regard,the stand-alone and genetic-code-independent peptide ligases constitute a new family of renegade peptide-bond makers.A prime example is butelase-1,an Asn/Asp(Asx)-specific ligase that structurally belongs to the asparaginyl endopeptidase family.Butelase-1 specifically recognizes a C-terminal Asx-containing tripeptide motif,Asn/Asp-Xaa-Yaa(Xaa and Yaa are any amino acids),to form a site-specific Asn-Xaa peptide bond either intramolecularly as cyclic proteins or intermolecularly as modified proteins.Our work in the past five years has validated that butelase-1 is a potent and versatile ligase.Here we review the advances in ligases,with a focus on butelase-1,and their applications in engineering bioactive peptides and precision protein modifications,antibody-drug conjugates,and live-cell labeling.展开更多
基金financial support from RGC Senior Research Fellow Scheme(SRFS2122-5S04),Research Grants Council of Hong KongCollaborative Research Fund(C5044-21GF)from the Research Grants Council of Hong Kong and National Excellent Young Scientists Fund(Hong Kong and Macao,82122002),National Natural Science Foundation of China+1 种基金support from National Natural Science Foundation of China(No.32071337)Shanghai Pujiang Program(20PJ1402600).
文摘Preservation of growth factor sensitivity and bioactivity(e.g.,bone morphogenetic protein-2(BMP-2))post-immobilization to tissue engineering scaffolds remains a great challenge.Here,we develop a stable and soft surface modification strategy to address this issue.BMP-2(a model growth factor)is covalently immobilized onto homogeneous poly(glycidyl methacrylate)(PGMA)polymer brushes which are grafted onto substrate surfaces(Au,quartz glass,silica wafer,or common biomaterials)via surface-initiated atom transfer radical polymerization.This surface modification method multiplies the functionalized interfacial area;it is simple,fast,gentle,and has little effect on the loaded protein owing to the cilia motility.The immobilized BMP-2(i-BMP-2)on the surface of homogeneous PGMA polymer brushes exhibits excellent bioactivity(-87%bioactivity of free BMP-2 in vitro and 20%-50%higher than scaffolds with free BMP-2 in vivo),with conformation and secondary structure well-preserved after covalent immobilization and ethanol sterilization.Moreover,the osteogenic activity of i-BMP-2 on the nanoline pattern(PGMA-poly(N-isopropylacrylamide))shows-110%bioactivity of free BMP-2.This is superior compared to conventional protein covalent immobilization strategies in terms of both bioactivity preservation and therapeutic efficacy.PGMA polymer brushes can be used to modify surfaces of different tissue-engineered scaffolds,which facilitates in situ immobilization of growth factors,and accelerates repair of a wide range of tissue types.
基金This research was supported by the National Science Founda-tion of China(21971141)"Chenguang Program 18CG01"supported by Shanghai Education Development Foundation and Shanghai Municipal Education.
文摘Recently,multifunctional polymers for protein conjugation have been facilely synthesized through the multicomponent reactions(MCRs).In this mini-review,current progress in the generation of multifuncti onal polymers with protei n-reactive groups via MCRs is summarized.These mult functional polymers react with model therapeutic proteins,forming multifunctional polymer-protein conjugates,which are prototypes of sophisticated theranostic agents and antibacterial vaccine candidates.In comparison with the traditional multi-step synthesis,the preparation of multifunctional polymers for protein conjugation through MCRs is straightforward and convenient.Due to these properties,MCRs have the potential to become a new general strategy to achieve polymer-protein conjugates for biological and medical applications.
基金Weare grateful for the financial support from the Natural Science Foundation of China(grant nos.21778009,21977010,and 22007033)National Key Research and Development Program“Synthetic Biology”Key Special Project of China(grant no.2018YFA0902504)+3 种基金China Postdoctoral Science Foundation(grant no.2021M690220)the Natural Science Foundation of Guangdong Province(grant nos.2020A1515010522,2020A1515010766,2019A1515110487,and 2019A151-5111184)the Foundation for Basic and Applied Research of Guangdong Province(grant no.2019A1515110489)and the Shenzhen Science and Technology Innovation Committee(grant nos.JCYJ20180507181527112,JCYJ-201805081522131455,and JCYJ20170817172023838).
文摘Over the past 20 years,great efforts have been invested in developing site-specific approaches to protein modification to dissect protein functions directly and accurately.Here,we report a proximitytriggered group transfer strategy from a sulfonium warhead to a Cysteine(Cys)residue of the target protein.With a guiding ligand,cargoes could be transferred selectively from a sulfonium center onto the Cys residue in the vicinity of their binding interface.The successful thalidomide transfer of sulfonium 1-X could be applied intracellularly for epidermal growth factor receptor degradation,highlighting the potential of group transfer strategy as a suite of chemical biology studies,including cell imaging,protein profiling,and protein degradation by simply employing different transferrable groups.
基金The National Natural Science Foundation of China (Nos. 21462023, 21778025)the Education Department of Jiangxi Province (No.150297)
文摘Artificial synthesis and site-specific modification of peptides and proteins has evolved into an indispensable tool for protein engineers and chemical biologists. Chemical and enzymatic approaches to peptide ligation are important alternatives of recombinant DNA technology for protein synthesis and modification. Although as old as that of chemical procedures, enzyme-mediated peptide ligation is far less developed than that of chemical counterpart due to the difficult availability of peptide ligase.Fortunately, this situation has been changed slowly with the fast development of biological techniques. In the past decades, several natural peptide ligases have been discovered. Protein engineering to improve the ligation efficiencies of the natural peptide ligase and to reverse the functionality of protease provide more powerful peptide ligases. In this review, the advances of enzyme-mediated peptide ligation and their application in protein synthesis and modification will be discussed.
基金supported by Academic Research Grant Tier 3(MOE2016-T3-1-003)from the Singapore Ministry of Education.
文摘Making peptide bonds is tightly controlled by genetic code and machinery which includes cofactors,ATP,and RNAs.In this regard,the stand-alone and genetic-code-independent peptide ligases constitute a new family of renegade peptide-bond makers.A prime example is butelase-1,an Asn/Asp(Asx)-specific ligase that structurally belongs to the asparaginyl endopeptidase family.Butelase-1 specifically recognizes a C-terminal Asx-containing tripeptide motif,Asn/Asp-Xaa-Yaa(Xaa and Yaa are any amino acids),to form a site-specific Asn-Xaa peptide bond either intramolecularly as cyclic proteins or intermolecularly as modified proteins.Our work in the past five years has validated that butelase-1 is a potent and versatile ligase.Here we review the advances in ligases,with a focus on butelase-1,and their applications in engineering bioactive peptides and precision protein modifications,antibody-drug conjugates,and live-cell labeling.
