0d8853a33f9b88a9c1d0645fcb58f092 pmc_32640506.pdf f67b1f3281220fbde35a1bee84066eb3ba0a6544 pmc_32640506.pdf 8fa6ff3093fbfb26f31ebac738723fae2c83ff12207a6fbd7b1fb45cc30ba5ae pmc_32640506.pdf Title: Magnetite Nanoparticles Functionalized with RNases against Intracellular Infection of Pseudomonas aeruginosa Subject: Current treatments against bacterial infections have severe limitations, mainly due to the emergence of resistance to conventional antibiotics. In the specific case of Pseudomonas aeruginosa strains, they have shown a number of resistance mechanisms to counter most antibiotics. Human secretory RNases from the RNase A superfamily are proteins involved in a wide variety of biological functions, including antimicrobial activity. The objective of this work was to explore the intracellular antimicrobial action of an RNase 3/1 hybrid protein that combines RNase 1 high catalytic and RNase 3 bactericidal activities. To achieve this, we immobilized the RNase 3/1 hybrid on Polyetheramine (PEA)-modified magnetite nanoparticles (MNPs). The obtained nanobioconjugates were tested in macrophage-derived THP-1 cells infected with Pseudomonas aeruginosa PAO1. The obtained results show high antimicrobial activity of the functionalized hybrid protein (MNP-RNase 3/1) against the intracellular growth of P. aeruginosa of the functionalized hybrid protein. Moreover, the immobilization of RNase 3/1 enhances its antimicrobial and cell-penetrating activities without generating any significant cell damage. Considering the observed antibacterial activity, the immobilization of the RNase A superfamily and derived proteins represents an innovative approach for the development of new strategies using nanoparticles to deliver antimicrobials that counteract P. aeruginosa intracellular infection. Keywords: Pseudomonas aeruginosa; ribonucleases; magnetite nanoparticles; antimicrobials Author: Nathaly Rangel-Muñoz, Alejandra Suarez-Arnedo, Raúl Anguita, Guillem Prats-Ejarque, Johann F. Osma, Carolina Muñoz-Camargo, Ester Boix, Juan C. Cruz and Vivian A. Salazar Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Mon Jul 6 07:10:45 2020 CEST ModDate: Mon Jul 6 07:10:45 2020 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 25 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4986554 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- EYSUOW+URWPalladioL-Ital Type 1 Custom yes yes no 153 0 WVPTUL+URWPalladioL-Bold Type 1 Custom yes yes no 154 0 BGVZXL+URWPalladioL-BoldItal Type 1 Custom yes yes no 155 0 QTLELQ+Pxbsy Type 1 Builtin yes yes no 156 0 ENLEMO+VnURWPalladioL-Bold Type 1 Custom yes yes no 157 0 HTIDNB+URWPalladioL-Roma Type 1 Custom yes yes no 158 0 EAGQMG+VnURWPalladioL Type 1 Custom yes yes no 159 0 DTNNLP+Pxsy Type 1 Builtin yes yes no 160 0 WDLIVJ+Rpxr Type 1 Builtin yes yes no 161 0 IAFAOG+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 169 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 190 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 191 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 192 0 YICMPF+Rpcxr Type 1 Builtin yes yes no 281 0 ZXDKIP+EURM10 Type 1 Builtin yes yes no 302 0 YWGLGW+Rpxi Type 1 Builtin yes yes no 303 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 359 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 360 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 361 0 FNTSBS+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 362 0 BDNPPA+PalatinoLinotype TrueType WinAnsi yes yes no 408 0 BDNPPA+PalatinoLinotype TrueType WinAnsi yes yes no 433 0 BDOBBL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 434 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 456 0 FNTSBS+Arial-BoldItalicMT CID TrueType Identity-H yes yes yes 457 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 458 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 459 0 FNTSBS+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 460 0 FNTSBS+Arial-BoldMT CID TrueType Identity-H yes yes yes 461 0 FNTSBS+SymbolMT,Bold CID TrueType Identity-H yes yes yes 462 0 FNTSBS+Calibri-Bold CID TrueType Identity-H yes yes yes 463 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 524 0 FNTSBS+Arial-BoldItalicMT CID TrueType Identity-H yes yes yes 525 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 526 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 527 0 FNTSBS+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 528 0 FNTSBS+Arial-BoldMT CID TrueType Identity-H yes yes yes 529 0 FNTSBS+SymbolMT,Bold CID TrueType Identity-H yes yes yes 530 0 FNTSBS+Calibri-Bold CID TrueType Identity-H yes yes yes 531 0 FNTSBS+ArialMT CID TrueType Identity-H yes yes yes 532 0 BDNPPA+PalatinoLinotype TrueType WinAnsi yes yes no 619 0 BDOBBL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 620 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 638 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 639 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 640 0 FNTSBS+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 641 0 BDNPPA+PalatinoLinotype TrueType WinAnsi yes yes no 678 0 BDOBBL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 679 0 NBHIAB+VnURWPalladioL-Italic Type 1 Custom yes yes no 781 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2020-11-18 06:23:17 CET RepresentationInformation: pmc_32640506.pdf ReportingModule: BYTESTREAM, Rel. 1.4 (2018-10-01) LastModified: 2020-08-17 03:36:27 CEST Size: 4986554 Format: bytestream Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/octet-stream Checksum: 84d024f5 Type: CRC32 Checksum: 0d8853a33f9b88a9c1d0645fcb58f092 Type: MD5 Checksum: f67b1f3281220fbde35a1bee84066eb3ba0a6544 Type: SHA-1