d42ea16f51de611ceb56b5c6b6e77160 ijms_a2018v19n3p678.pdf 59eb5279c458ef27065c1254be45c133008bc193 ijms_a2018v19n3p678.pdf 92300ed88da442eaaeb37ceeb26bf6f2f7c3a9730ca5fd124085895e560fb1f5 ijms_a2018v19n3p678.pdf Syntax Error: Couldn't find trailer dictionary Syntax Error: Invalid XRef entry Syntax Error: Top-level pages object is wrong type (null) Syntax Error: Top-level pages object is wrong type (null) Syntax Error: Top-level pages object is wrong type (null) Syntax Error: Top-level pages object is wrong type (null) Title: Biochemical and MALDI-TOF Mass Spectrometric Characterization of a Novel Native and Recombinant Cystine Knot Miniprotein from Solanum tuberosum subsp. andigenum cv. Churqueña Subject: Cystine-knot miniproteins (CKMPs) are an intriguing group of cysteine-rich molecules that combine the characteristics of proteins and peptides. Typically, CKMPs are fewer than 50 residues in length and share a characteristic knotted scaffold characterized by the presence of three intramolecular disulfide bonds that form the singular knotted structure. The knot scaffold confers on these proteins remarkable chemical, thermal, and proteolytic stability. Recently, CKMPs have emerged as a novel class of natural molecules with interesting pharmacological properties. In the present work, a novel cystine-knot metallocarboxypeptidase inhibitor (chuPCI) was isolated from tubers of Solanum tuberosum, subsp. andigenum cv. Churqueña. Our results demonstrated that chuPCI is a member of the A/B-type family of metallocarboxypeptidases inhibitors. chuPCI was expressed and characterized by a combination of biochemical and mass spectrometric techniques. Direct comparison of the MALDI-TOF mass spectra for the native and recombinant molecules allowed us to confirm the presence of four different forms of chuPCI in the tubers. The majority of such forms have a molecular weight of 4309 Da and contain a cyclized Gln in the N-terminus. The other three forms are derived from N-terminal and/or C-terminal proteolytic cleavages. Taken together, our results contribute to increase the current repertoire of natural CKMPs. Keywords: cystine-knot miniproteins; carboxypeptidase inhibitor; plant inhibitor; Solanum tuberosum; protease; Andean potatoes Author: Juliana Cotabarren, Mariana Edith Tellechea, Sebastián Martín Tanco, Julia Lorenzo, Javier Garcia-Pardo, Francesc Xavier Avilés and Walter David Obregón Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.15; modified using iText® 7.0.1 ©2000-2016 iText Group NV (AGPL-version) CreationDate: Wed Feb 28 09:42:37 2018 CET ModDate: Wed Feb 28 09:45:49 2018 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 0 x 0 pts Page rot: 0 File size: 3365508 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JYPLJJ+VnURWPalladioL-Bold Type 1 Custom yes yes no 127 0 OXBKYU+TeXGyreAdventor-Bold Type 1 Custom yes yes no 128 0 MWHIBA+VnURWPalladioL Type 1 Custom yes yes no 130 0 EKNHNV+URWPalladioL-Ital Type 1 Custom yes yes no 124 0 DONVLT+URWPalladioL-Bold Type 1 Custom yes yes no 125 0 JCTRLH+URWPalladioL-BoldItal Type 1 Custom yes yes no 126 0 NSLIWV+URWPalladioL-Roma Type 1 Custom yes yes no 129 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 136 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 135 0 ALMJEC+EURM10 Type 1 Builtin yes yes no 192 0 LLMNHJ+CMSY10 Type 1 Builtin yes yes no 235 0 MEGPOB+PalatinoLinotype CID TrueType Identity-H yes yes yes 254 0 MEHBEB+TimesNewRoman TrueType WinAnsi yes yes no 255 0 MEGPPB+PalatinoLinotype TrueType WinAnsi yes yes no 256 0 MEHAGA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 257 0 MEHAHC+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 258 0 MEGPOB+PalatinoLinotype CID TrueType Identity-H yes yes yes 297 0 MEHBEB+TimesNewRoman TrueType WinAnsi yes yes no 298 0 MEGPPB+PalatinoLinotype TrueType WinAnsi yes yes no 299 0 MEHAGA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 300 0 MEHAHC+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 301 0 MEHBEB+TimesNewRoman TrueType WinAnsi yes yes no 353 0 MEGPPB+PalatinoLinotype TrueType WinAnsi yes yes no 354 0 MEHAGA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 355 0 MEHAHC+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 356 0 MEGPOB+PalatinoLinotype CID TrueType Identity-H yes yes yes 383 0 MEHBEB+TimesNewRoman TrueType WinAnsi yes yes no 384 0 MEGPPB+PalatinoLinotype TrueType WinAnsi yes yes no 385 0 MEHAGA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 386 0 MEHAHC+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 387 0 MEHBEB+TimesNewRoman TrueType WinAnsi yes yes no 441 0 MEGPPB+PalatinoLinotype TrueType WinAnsi yes yes no 442 0 MEHAGA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 443 0 MEHAHC+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 444 0 CZNUER+CMEX10 Type 1 Builtin yes yes no 480 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 478 0 SOSTRQ+CMR10 Type 1 Builtin yes yes no 476 0 Jhove (Rel. 1.6, 2011-01-04) Date: 2019-02-02 04:48:15 CET RepresentationInformation: ijms_a2018v19n3p678.pdf ReportingModule: BYTESTREAM, Rel. 1.3 (2007-04-10) LastModified: 2019-01-29 12:05:26 CET Size: 3365508 Format: bytestream Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/octet-stream Checksum: 5b6d5d7d Type: CRC32 Checksum: d42ea16f51de611ceb56b5c6b6e77160 Type: MD5 Checksum: 59eb5279c458ef27065c1254be45c133008bc193 Type: SHA-1