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SKOV-3 cell imaging by paramagnetic particles labeled with hairpin Cell-penetrating peptides

Chinese Medical Journal 2011年第1期124卷 111-117页

作者： ZHAI Xiao-hui LIU Min GUO Xiao-juan WANG Si-cen ZHANG Hong-xia GUO You-min Imaging Center the First Affiliated Hospital of Medical School Xi'an Jiao Tong University Xi'an 710004 Shannxi China Department of Radiology Beijing Chao Yang Hospital Capital Medical University Beijing 100020 China Department of Pharmacy Medical School Xi'an Jiao TongUniversity Xi'an Shannxi 710061 China

Background The hairpin Cell-penetrating peptides （hCPPs） demonstrate an interesting characteristic of conditioned activation by molecules. We hypothesized that hCPPs have the potential to selectively deliver a paramagnetic gadolinium probe into the matrix metalloproteinase 2 （MMP-2） positive human ovary adenocarcinoma cell lines, SKOV-3. Methods hCPPs were synthesized and labeled with 1,4,7,10-tetraazacyclododecane-N,N＇,N＂,N＂＇-tetraacetic acid gadolinium （111）（Gd-DOTA） and fluorescein isothiocyanate （FITC） by f-moc strategy using a standard solid phase peptide synthesis protocol. MMP-2 expression and activity were demonstrated by reverse transcriptase polymerase chain reaction （RT-PCR） and zymography. Internalization and location of hCPPs in SKOV-3 cells were observed by fluorescein imaging and flow cytometery. Selective delivery of Gd-DOTA in SKOV-3 cells was observed by magnetic resonance imaging （MRI） and transmission electron microscopy （TEM）. Results The uptake of hCPPs by SKOV-3 cells depended on the activity of MMP-2. T1WI signals of SKOV-3 cells treated with Gd-DOTA-hCPPs suggested the uptake of Gd-DOTA-hCPPs increased in a time- （r=0.990, P 〈0.01） and concentration-dependent manner （r=0.964, P 〈0.001）, but was inhibited by a MMP-2 inhibitor. Electron-dense particles observed in the cytoplasm and nucleus by transmission electron microscopy proved the intracellular penetration of gadolinium. Conclusions hCPPs can be used as an effective vector for an MRI molecular probe to assess the activity of MMP-2.

关键词： Cell-penetrating peptides matrix metalloproteinase-2 molecular imaging,＂ magnetic resonance imaging

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Translocation of cell penetrating peptides

Translocation of Cell Penetrating Peptides

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中国化学会第29届学术年会

作者： Xinli Gao Hongsong Xianren Zhang State Key Laboratory of Organic-Inorganic Composites Beijing University of Chemical Technology

In real cell environment,as we all know,ionic charge imbalance is ever present,thus forming electrostatic potential cross the membrane.cellular processes,including the delivery of drug,polymer,micro-and macro-molecule,may rely on transmembrane potential.A number of experimental studies demonstrate that Cell-penetrating peptides（CPPs）can enter cells in an energy-independent way,which is known as direct translocation.The entry is complementary to endocytosis and featured by the rapid uptake,making CPPs as a fascinating intracellular deliver agent.The ability of CPPs that can carry drugs into cells,however,the mechanism of CPP penetrating is still unknown.Here,molecular dynamics simulation method is applied to show that transmembrane potential induced by ion imbalance plays an important role for the translocation of CPPs.Furthermore,during the translocation process,a nanoscale hydrophilic hole is found in the lipid membrane,which can facilitate CPPs transporting into cell through membrane.This findings not only interpret the mechanism of CPPs penetrating across membrane,but also demonstrate the importantce of transmembrane potential in cell activity.

关键词： ion imbalance Cell-penetrating peptides lipid membrane tranmembrane potential nanoscale hole

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Transmembrane Potential enhances Direct Translocation of Cell-penetrating peptides

Transmembrane Potential enhances Direct Translocation of Cel...

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中国化学会2016年软物质理论计算与模拟会议

作者： Xinli Gao Song Hong Tongtao Yue Zhiping Liu Jure Dobnikar Xianren Zhang State Key Laboratory of Organic-Inorganic Composites Beijing University of Chemical Technology State Key Laboratory of Heavy Oil Processing Center for Bioengineering and BiotechnologyChina University of Petroleum (East China) International Research Center for Soft Matter Beijing University of Chemical Technology

Cell-penetrating peptides（CPPs） are shown experimentally to be capable of entering cells in an energy-independent pathway of direct translocation,which is complementary to endocytosis and featured by the rapid uptake,making CPPs a fascinating drug delivery agent.The mechanisms underlying the direct penetration of CPPs,however,are still open challenges.In this work,both direct molecular dynamics simulations and free energy calculations demonstrate that the transmembrane potential due to the ion concentration difference between the intracellular and extracellular regions and local membrane potential enhancement due to CPP adsorption play essential roles in the direct translocation of CPPs.In particular,our simulations show that the local membrane potential enhancement caused by highly charged CPP structures and by high CPP density on the membranes cooperatively promote the formation of a membrane pore,through which the peptides could penetrate into the membrane hole from one end with a chain-like structure.These findings not only interpret the mechanism of CPP penetration across membrane,but also demonstrate the importance of transmembrane potential for many cell activities.

关键词： Cell-penetrating peptides membrane membrane potential translocation

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cell-penetrating riboflavin conjugate for antitumor photodynamic therapy

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Chinese Chemical Letters 2022年第9期33卷 4339-4344页

作者： Chunlei Wu Yanyan Li Zhehong Cheng Pengxin Wang Zhilong Ma Ke Liu Yulian Cheng Yimin Zhou Xian Lin Ximing Shao Yong Yang Hongchang Li Lijing Fang Guangdong Key Laboratory of Nanomedicine Institute of Biomedicine and BiotechnologyShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhen 518055China University of Chinese Academy of Sciences Beijing 100049China Department of Biomedical Engineering Southern University of Science and Technology(SUSTech)Shenzhen 518055China

Riboflavin(RF,vitamin B2)is an essential vitamin and has been considered as a promising natural photosensitizer for photodynamic therapy(PDT).However,further exploration of RF in antitumor application was limited by its poor cellular uptake.In this study,using Cell-penetrating peptides Arg8,(Cha-Arg)3 and small molecule triphenylphosphine(TPP)as delivery compounds,three RF conjugates were prepared to increase the accumulation of RF in cells,termed as Arg8-RF,(Cha-Arg)3-RF and TPP-RF,respectively.Compared with TPP-RF and Arg8-RF,(Cha-Arg)3-RF exhibited better cell internalization and stronger cytotoxicity against HeLa cells upon exposure to blue light.Further researches proved that(Cha-Arg)3-RF generated reactive oxygen species(ROS)under irradiation,which could indiscriminately destroy endogenous proteins and mitochondria,ultimately inducing cell death.This work provides a new approach to explore RF as a natural photosensitizer for antitumor photodynamic therapy.

关键词： Riboflavin Antitumor Photodynamic therapy Cell-penetrating peptides

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peptides-based vaccine against SARS-nCoV-2 antigenic fragmented synthetic epitopes recognized by T cell and β-cell initiation of specific antibodies tofightthe infection

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Bio-Design and Manufacturing 2021年第3期4卷 490-505页

作者： Zainularifeen Abduljaleel Faisal A.Al-Allaf Syed A.Aziz Department of Medical Genetics Faculty of MedicineUmm Al-Qura UniversityP.O.Box715Mecca21955Kingdom of Saudi Arabia Science and Technology Unit UmmAl-Qura UniversityP.O.Box 715Mecca21955Kingdom of SaudiArabia The Regional Laboratory Molecular Diagnostics UnitDepartment of Molecular BiologyMinistry of Health(MOH)P.O.Box 6251MeccaKingdom of SaudiArabia Department of Pathology and Lab Medicine University of Ottawa451Smyth RoadOttawaONK1H8M5Canada

The World Health Organization has declared the rapidly spreading coronavirus to be a global pandemic.The FDA is yet to approve a vaccine for human novel coronavirus.Here,we developed a peptide-based vaccine and used high-throughput screening by molecular dynamics simulation to identify T-cell-and p-cell-recognized epitopes for producing specific antibod-ies against SARS-nCoV-2.We construct~12 P'antigenic epitope peptides to develop a more effective vaccine and identify specific antibodies.These epitope peptides selectively presented the best antigen presentation scores for both human pMHC class I and II alleles to develop a strong binding affinity.All antigens identified of SARS-nCoV-2 different proteins by each attached specific~1-7 L linker adaptor were used to construct a broad single peripheral peptide vaccine.It is expected to be highly antigenic with a minimum allergic effect.As a result of these exciting outcomes,expressing a vaccine using the intimated peptide was highly promising and positive to be highly proposed as epitope-based peptide vaccine of specific antibody against SARS-nCoV-2 by initiating T cells and β-cells.An in vitro study for the proposed peptide-based vaccine is.mostly recommended.Further clinical trials are required to check the efficacy of this vaccine.

关键词： COVID-19 SARS-nCoV-2 Peptide-based vaccine Cell-penetrating peptides Lipid membrane Vaccine efficacy

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Study on the Penetrability of PEP-1-P27mt for cell Membranes in Vitro

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2007年第3期27卷 225-229页

作者：严世荣朱明磊邱方城王立林屈伸 Department of Clinical Laboratory Shiyan People's Hospital Affiliated to Yunyang Medical College Department of Biochemistry and Molecular Biology School of Basic Medical Sciences Tongji Medical College Huazhong University of Science and Technology

Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, pro- karyotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS trans- formed and induced with IPTG to highly express fusion protein PEP-1-P27mt. Fusion protein with an N-terminal His-tag could be purified by Ni2+-resin affinity chromatography and identified by SDS-PAGE and Western blotting. Cultured EC9706 cells treated with PEP-1-P27mt revealed that PEP-1-P27mt was transduced into cells after 15 min and reached maximal intracellular concentra- tions in 2 h. PEP-1-P27mt of 1 μmol/L final concentration could most strongly suppress the growth. It was suggested that PEP-1 can carry P27mt across membrane, which provides a new biological pro- tocol for using cyclin dependent kinase inhibitors p27mt in suppressing the growth of tumor cells.

关键词： Cell-penetrating peptides PEP-1 p27mt pET15b-pep-1-p27mt recombinant

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