c689f5ced5a6f276a5a1583785d63407 pmc_28821821.pdf 235f7d45afd65dc53c3adaf217f7f1ab944afb67 pmc_28821821.pdf 3edcb5fcbf154ea3962d056cd9c7a663c9d012f1f5ed510b4380d1f2ca22fa1a pmc_28821821.pdf Title: The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase: A mutagenesis analysis Subject: Scientific Reports, doi:10.1038/s41598-017-09360-5 Keywords: Author: Cristina Molero Creator: Springer Producer: iText® 5.3.5 ©2000-2012 1T3XT BVBA (AGPL-version) CreationDate: Fri Aug 11 13:00:01 2017 CEST ModDate: Mon Aug 14 17:08:56 2017 CEST Tagged: no UserProperties: no Suspects: no Form: AcroForm JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 782.362 pts Page rot: 0 File size: 3013162 bytes Optimized: no PDF version: 1.4 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- RSUHPK+Corbel CID TrueType Identity-H yes yes yes 459 0 BDCWKH+Corbel-BoldItalic Type 1C WinAnsi yes yes yes 318 0 MinionPro-It Type 1C Custom yes no yes 63 0 MinionPro-Regular Type 1C Custom yes no yes 68 0 HUISPO+Corbel-Bold2 Type 1C WinAnsi yes yes yes 201 0 EuclidSymbol Type 1C Custom yes no yes 62 0 LXWXQE+Corbel2 Type 1C WinAnsi yes yes yes 66 0 TLUORG+Corbel-Bold CID TrueType Identity-H yes yes yes 67 0 EuclidSymbol-Bold Type 1C Custom yes no yes 460 0 OXWQUQ+TimesNewRomanPSMT Type 1C WinAnsi yes yes yes 461 0 HUISPO+Corbel-Bold2 Type 1C WinAnsi yes yes yes 356 0 LXWXQE+Corbel2 Type 1C WinAnsi yes yes yes 357 0 RSUHPK+Corbel CID TrueType Identity-H yes yes yes 504 0 MyriadPro-It Type 1C WinAnsi yes no yes 359 0 MinionPro-Bold Type 1C WinAnsi yes no yes 64 0 EuclidSymbol-Italic Type 1C Custom yes no yes 65 0 LXWXQE+Corbel2 Type 1C WinAnsi yes yes yes 352 0 RSUHPK+Corbel CID TrueType Identity-H yes yes yes 358 0 HUISPO+Corbel-Bold2 Type 1C WinAnsi yes yes yes 366 0 RSUHPK+Corbel CID TrueType Identity-H yes yes yes 365 0 ZTFZSU+STIXGeneral-Regular Type 1C Custom yes yes yes 202 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-02-08 02:46:22 CET RepresentationInformation: pmc_28821821.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2018-02-08 07:51:42 CET Size: 3013162 Format: PDF Version: 1.4 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 570 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseOutlines Language: EN Outlines: Item: Title: The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase: A mutagenesis analysis Children: Item: Title: Results Children: Item: Title: Screen of a random-mutagenesis library for deregulated versions of Ppz1. Children: Item: Title: Phenotypic analysis of mutated alleles of PPZ1. Item: Title: Determination of the capacity Hal3 to inhibit the Ppz1 variants. Item: Title: Ppz1 variants retain the ability to bind Hal3. Item: Title: Characterization of additional PPZ1 alleles. Item: Title: Mapping the mutations in Ppz1 and PP1c sequences. Item: Title: Discussion Item: Title: Methods Children: Item: Title: Yeast strains and growth conditions. Item: Title: Recombinant DNA techniques and plasmids. Item: Title: Random PCR Mutagenesis and screen for Ppz1 deregulated versions. Item: Title: Yeast protein extraction and immunoblot analysis. Item: Title: Expression of Ppz1 mutated versions in E. coli. Item: Title: Assay of protein phosphatase activity. Item: Title: Detection of protein-protein interactions. Item: Title: Phenotypic analysis of yeast cells. Item: Title: Data availability. Item: Title: Acknowledgements Item: Title: Figure 1 Growth on caffeine plates of selected Ppz1 clones carrying single amino acid mutations. Item: Title: Figure 2 Tolerance to Li+ cations conferred by the diverse variants of the Ppz1 phosphatase. Item: Title: Figure 3 Ppz1 variants show differences in the pattern of in vitro inhibition by Hal3. Item: Title: Figure 4 Interaction of Hal3 with the different Ppz1 variants. Item: Title: Figure 5 Phenotypic analysis and expression levels of selected Ppz1 versions carrying two amino acid changes. Item: Title: Figure 6 Hal3 binding capacity and inhibition profile of Ppz1 variants carrying two aminoacid mutations. Item: Title: Figure 7 Alignment of the catalytic domains of Ppz1 from S. Item: Title: Figure 8 Mapping mutations and functional elements on a structural model of the C-terminal catalytic moiety of ScPpz1. Item: Title: Table 1 Mutations affecting the catalytic domain of Ppz1 that result in single amino acid change. Item: Title: Table 2 Mutations affecting the catalytic domain of Ppz1 that result in changes in two amino acids. Item: Title: Table 3 Yeast strains used in this work. Info: Title: The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase: A mutagenesis analysis Author: Cristina Molero Subject: Scientific Reports, doi:10.1038/s41598-017-09360-5 Keywords: Creator: Springer Producer: iText® 5.3.5 ©2000-2012 1T3XT BVBA (AGPL-version) CreationDate: Fri Aug 11 13:30:01 CEST 2017 ModDate: Mon Aug 14 17:08:56 CEST 2017 ID: 0xec3256b85b1c6eae406c041b30d69d09, 0xecc94b182e5f43e563346685b10be4df Filters: FilterPipeline: FlateDecode FilterPipeline: DCTDecode Images: Image: NisoImageMetadata: FormatName: image/png ImageWidth: 1100 ImageHeight: 90 ColorSpace: black is zero BitsPerSample: 1 BitsPerSampleUnit: integer Filter: FlateDecode Intent: RelativeColorimetric Name: X Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1500 ImageHeight: 1941 ColorSpace: black is zero BitsPerSample: 8 BitsPerSampleUnit: integer Intent: RelativeColorimetric Name: X Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1650 ImageHeight: 1182 ColorSpace: black is zero BitsPerSample: 8 BitsPerSampleUnit: integer Intent: RelativeColorimetric Name: X Image: NisoImageMetadata: FormatName: image/png ImageWidth: 1900 ImageHeight: 901 ColorSpace: black is zero BitsPerSample: 8 BitsPerSampleUnit: integer Filter: FlateDecode Intent: RelativeColorimetric Name: X Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1650 ImageHeight: 1231 ColorSpace: black is zero BitsPerSample: 8 BitsPerSampleUnit: integer Intent: RelativeColorimetric Name: X Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1232 ImageHeight: 2300 ColorSpace: black is zero BitsPerSample: 8 BitsPerSampleUnit: integer Intent: RelativeColorimetric Name: X Image: NisoImageMetadata: FormatName: image/png ImageWidth: 1650 ImageHeight: 741 ColorSpace: black is zero BitsPerSample: 8 BitsPerSampleUnit: integer Filter: FlateDecode Intent: RelativeColorimetric Name: X Image: NisoImageMetadata: FormatName: image/png ImageWidth: 1900 ImageHeight: 2150 BitsPerSample: 8 BitsPerSampleUnit: integer Filter: FlateDecode Intent: RelativeColorimetric Name: X Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1650 ImageHeight: 956 BitsPerSample: 8 BitsPerSampleUnit: integer Intent: RelativeColorimetric Name: X Fonts: Type0: Font: BaseFont: TLUORG+Corbel-Bold Encoding: Identity-H ToUnicode: true Font: BaseFont: RSUHPK+Corbel Encoding: Identity-H ToUnicode: true Type1: Font: BaseFont: MinionPro-Bold FirstChar: 0 LastChar: 255 FontDescriptor: FontName: MinionPro-Bold Flags: Serif, Nonsymbolic FontBBox: -319, -360, 1684, 1024 FontFile3: true Encoding: WinAnsiEncoding ToUnicode: true Font: BaseFont: EuclidSymbol-Italic FirstChar: 0 LastChar: 255 FontDescriptor: FontName: EuclidSymbol-Italic Flags: Symbolic, Italic FontBBox: -166, -276, 1162, 950 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true Font: BaseFont: LXWXQE+Corbel2 FontSubset: true FirstChar: 0 LastChar: 255 FontDescriptor: FontName: LXWXQE+Corbel2 Flags: Nonsymbolic FontBBox: -553, -239, 1110, 952 FontFile3: true Encoding: WinAnsiEncoding ToUnicode: true Font: BaseFont: MinionPro-Regular FirstChar: 0 LastChar: 255 FontDescriptor: FontName: MinionPro-Regular Flags: Serif, Nonsymbolic FontBBox: -290, -360, 1684, 989 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true Font: BaseFont: HUISPO+Corbel-Bold2 FontSubset: true FirstChar: 0 LastChar: 255 FontDescriptor: FontName: HUISPO+Corbel-Bold2 Flags: Nonsymbolic FontBBox: -556, -239, 1155, 952 FontFile3: true Encoding: WinAnsiEncoding ToUnicode: true Font: BaseFont: ZTFZSU+STIXGeneral-Regular FontSubset: true FirstChar: 29 LastChar: 181 FontDescriptor: FontName: ZTFZSU+STIXGeneral-Regular Flags: Serif, Nonsymbolic FontBBox: -970, -443, 2000, 1023 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true Font: BaseFont: EuclidSymbol-Bold FirstChar: 0 LastChar: 255 FontDescriptor: FontName: EuclidSymbol-Bold Flags: Symbolic FontBBox: -186, -308, 1101, 960 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true Font: BaseFont: OXWQUQ+TimesNewRomanPSMT FontSubset: true FirstChar: 32 LastChar: 121 FontDescriptor: FontName: OXWQUQ+TimesNewRomanPSMT Flags: Serif, Nonsymbolic FontBBox: -568, -307, 2000, 1007 FontFile3: true Encoding: WinAnsiEncoding ToUnicode: true Font: BaseFont: BDCWKH+Corbel-BoldItalic FontSubset: true FirstChar: 0 LastChar: 255 FontDescriptor: FontName: BDCWKH+Corbel-BoldItalic Flags: Nonsymbolic, Italic FontBBox: -494, -239, 1194, 952 FontFile3: true Encoding: WinAnsiEncoding ToUnicode: true Font: BaseFont: EuclidSymbol FirstChar: 0 LastChar: 255 FontDescriptor: FontName: EuclidSymbol Flags: Symbolic FontBBox: -166, -253, 1090, 950 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true Font: BaseFont: MinionPro-It FirstChar: 0 LastChar: 255 FontDescriptor: FontName: MinionPro-It Flags: Serif, Nonsymbolic, Italic FontBBox: -201, -360, 1684, 1002 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true CIDFontType2: Font: BaseFont: TLUORG+Corbel-Bold CIDSystemInfo: Registry: Adobe Registry: Identity Supplement: 0 FontDescriptor: FontName: TLUORG+Corbel-Bold Flags: Symbolic FontBBox: -556, -239, 1155, 952 FontFile2: true Font: BaseFont: RSUHPK+Corbel CIDSystemInfo: Registry: Adobe Registry: Identity Supplement: 0 FontDescriptor: FontName: RSUHPK+Corbel Flags: Symbolic FontBBox: -553, -239, 1110, 952 FontFile2: true XMP: application/pdf doi:10.1038/s41598-017-09360-5 Springer US Scientific Reports, doi:10.1038/s41598-017-09360-5 The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase: A mutagenesis analysis Cristina Molero Carlos Casado Joaquamp#x000ED;n Ariamp#x000F1;o 10.1038/s41598-017-09360-5 http://dx.doi.org/10.1038/s41598-017-09360-5 journal Scientific Reports © 0, The Author(s) 2045-2322 10.1038/s41598-017-09360-5 Springer 2017-08-14T17:08:56+02:00 2017-08-11T16:30:01+05:30 2017-08-14T17:08:56+02:00 True iText® 5.3.5 ©2000-2012 1T3XT BVBA (AGPL-version) VoR uuid:da5e0a16-35ec-42af-8a2a-31ec04fa1bbe uuid:9bbb668a-8f3e-4bf2-adeb-f23938a242d6 default 1 converted uuid:b42ef330-3740-43fc-87e3-eb30b17a4b0c converted to PDF/A-2b pdfToolbox 2017-08-11T16:30:43+05:30 2 B http://ns.adobe.com/xap/1.0/mm/ xmpMM XMP Media Management Schema internal UUID based identifier for specific incarnation of a document InstanceID URI internal The common identifier for all versions and renditions of a document. OriginalDocumentID URI http://ns.adobe.com/pdf/1.3/ pdf Adobe PDF Schema internal A name object indicating whether the document has been modified to include trapping information Trapped Text http://ns.adobe.com/pdfx/1.3/ pdfx pdfx internal ID of PDF/X standard GTS_PDFXVersion Text internal Conformance level of PDF/X standard GTS_PDFXConformance Text internal Company creating the PDF Company Text internal Date when document was last modified SourceModified Text internal Mirrors crossmark:DOI doi Text http://www.aiim.org/pdfa/ns/id/ pdfaid PDF/A ID Schema internal Part of PDF/A standard part Integer internal Amendment of PDF/A standard amd Text internal Conformance level of PDF/A standard conformance Text http://prismstandard.org/namespaces/basic/2.0/ prism Prism external The aggregation type specifies the unit of aggregation for a content collection. Comment PRISM recommends that the PRISM Aggregation Type Controlled Vocabulary be used to provide values for this element. Note: PRISM recommends against the use of the #other value currently allowed in this controlled vocabulary. In lieu of using #other please reach out to the PRISM group at info@prismstandard.org to request addition of your term to the Aggregation Type Controlled Vocabulary. aggregationType Text external Copyright copyright Text external The Digital Object Identifier for the article. The DOI may also be used as the dc:identifier. If used as a dc:identifier, the URI form should be captured, and the bare identifier should also be captured using prism:doi. If an alternate unique identifier is used as the required dc:identifier, then the DOI should be specified as a bare identifier within prism:doi only. If the URL associated with a DOI is to be specified, then prism:url may be used in conjunction with prism:doi in order to provide the service endpoint (i.e. the URL). doi Text external ISSN for an electronic version of the issue in which the resource occurs. Permits publishers to include a second ISSN, identifying an electronic version of the issue in which the resource occurs (therefore e(lectronic)Issn. If used, prism:eIssn MUST contain the ISSN of the electronic version. See prism:issn. issn Text external Title of the magazine, or other publication, in which a resource was/will be published. Typically this will be used to provide the name of the magazine an article appeared in as metadata for the article, along with information such as the article title, the publisher, volume, number, and cover date. Note: Publication name can be used to differentiate between a print magazine and the online version if the names are different such as “magazine” and “magazine.com.” publicationName Text external This element provides the url for an article or unit of content. The attribute platform is optionally allowed for situations in which multiple URLs must be specified. PRISM recommends that a subset of the PCV platform values, namely “mobile” and “web”, be used in conjunction with this element. NOTE: PRISM recommends against the use of the #other value allowed in the PRISM Platform controlled vocabulary. In lieu of using #other please reach out to the PRISM group at prism-wg@yahoogroups.com to request addition of your term to the Platform Controlled Vocabulary. url Text http://www.niso.org/schemas/jav/1.0/ jav NISO external Values for Journal Article Version are one of the following: AO = Author’s Original SMUR = Submitted Manuscript Under Review AM = Accepted Manuscript P = Proof VoR = Version of Record CVoR = Corrected Version of Record EVoR = Enhanced Version of Record journal_article_version Closed Choice of Text Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 458, 304, 460, 292 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 174, 284, 176, 272 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 179, 284, 182, 272 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 472, 284, 475, 272 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 477, 284, 480, 272 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 365, 274, 368, 262 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 359, 264, 361, 252 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 364, 264, 367, 252 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 369, 264, 372, 252 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 457, 264, 460, 252 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 187, 254, 190, 242 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 243, 244, 249, 232 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 253, 224, 259, 212 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 414, 224, 420, 212 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 190, 204, 196, 192 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 458, 184, 464, 172 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 421, 164, 427, 152 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 373, 144, 379, 132 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 382, 144, 387, 132 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 483, 114, 489, 102 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 492, 114, 498, 102 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 500, 114, 506, 102 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 509, 114, 515, 102 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 428, 53, 508, 43 Flags: Print AppearanceDictionary: true Thumb: true Page: Sequence: 2 Annotations: Annotation: Subtype: Link Rect: 255, 535, 261, 523 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 529, 525, 534, 513 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 538, 525, 544, 513 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 234, 515, 239, 503 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 516, 515, 519, 503 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 522, 515, 527, 503 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 530, 515, 536, 503 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 280, 465, 286, 453 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 288, 465, 294, 453 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 295, 455, 301, 443 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 293, 435, 298, 423 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 539, 425, 545, 413 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 362, 395, 368, 383 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 202, 385, 208, 373 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 210, 385, 216, 373 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 330, 375, 336, 363 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 237, 226, 242, 213 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 401, 176, 406, 163 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 319, 156, 323, 143 Flags: Print AppearanceDictionary: true Destination: 3 Annotation: Subtype: Link Rect: 526, 114, 531, 102 Flags: Print AppearanceDictionary: true Destination: 69 Annotation: Subtype: Link Rect: 433, 84, 438, 72 Flags: Print AppearanceDictionary: true Destination: 3 Annotation: Subtype: Link Rect: 395, 54, 399, 42 Flags: Print AppearanceDictionary: true Thumb: true Page: Sequence: 3 Annotations: Annotation: Subtype: Link Rect: 494, 128, 502, 116 Flags: Print AppearanceDictionary: true Destination: 84 Annotation: Subtype: Link Rect: 549, 78, 552, 66 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 217, 68, 225, 56 Flags: Print AppearanceDictionary: true Destination: 84 Thumb: true Page: Sequence: 4 Annotations: Annotation: Subtype: Link Rect: 545, 307, 549, 295 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 380, 287, 384, 275 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 413, 237, 419, 225 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 421, 237, 427, 225 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 520, 227, 524, 215 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 171, 93, 176, 80 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 296, 63, 300, 50 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 493, 431, 497, 419 Flags: Print AppearanceDictionary: true Destination: 69 Thumb: true Page: Sequence: 5 Annotations: Annotation: Subtype: Link Rect: 453, 390, 458, 378 Flags: Print AppearanceDictionary: true Destination: 110 Annotation: Subtype: Link Rect: 550, 350, 554, 338 Flags: Print AppearanceDictionary: true Destination: 69 Annotation: Subtype: Link Rect: 171, 340, 175, 328 Flags: Print AppearanceDictionary: true Destination: 84 Annotation: Subtype: Link Rect: 373, 330, 379, 318 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 522, 300, 528, 288 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 530, 300, 536, 288 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 538, 300, 544, 288 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 547, 300, 552, 288 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 274, 237, 282, 225 Flags: Print AppearanceDictionary: true Destination: 143 Annotation: Subtype: Link Rect: 279, 177, 288, 165 Flags: Print AppearanceDictionary: true Destination: 143 Annotation: Subtype: Link Rect: 472, 134, 477, 122 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 209, 114, 214, 102 Flags: Print AppearanceDictionary: true Destination: 143 Annotation: Subtype: Link Rect: 323, 94, 328, 82 Flags: Print AppearanceDictionary: true Thumb: true Page: Sequence: 6 Annotations: Annotation: Subtype: Link Rect: 282, 106, 313, 94 Flags: Print AppearanceDictionary: true Destination: 160 Annotation: Subtype: Link Rect: 329, 86, 334, 74 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 430, 66, 434, 54 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 351, 57, 355, 44 Flags: Print AppearanceDictionary: true Thumb: true Page: Sequence: 7 Annotations: Annotation: Subtype: Link Rect: 270, 94, 279, 82 Flags: Print AppearanceDictionary: true Destination: 160 Annotation: Subtype: Link Rect: 481, 84, 490, 72 Flags: Print AppearanceDictionary: true Destination: 177 Annotation: Subtype: Link Rect: 341, 154, 345, 142 Flags: Print AppearanceDictionary: true Destination: 69 Annotation: Subtype: Link Rect: 362, 154, 367, 142 Flags: Print AppearanceDictionary: true Destination: 84 Annotation: Subtype: Link Rect: 304, 144, 313, 132 Flags: Print AppearanceDictionary: true Destination: 84 Thumb: true Page: Sequence: 8 Annotations: Annotation: Subtype: Link Rect: 181, 451, 189, 439 Flags: Print AppearanceDictionary: true Destination: 177 Annotation: Subtype: Link Rect: 545, 307, 549, 295 Flags: Print AppearanceDictionary: true Destination: 204 Annotation: Subtype: Link Rect: 304, 297, 310, 285 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 277, 227, 282, 215 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 548, 197, 552, 185 Flags: Print AppearanceDictionary: true Destination: 219 Annotation: Subtype: Link Rect: 546, 115, 552, 103 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 454, 105, 460, 93 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 227, 528, 236, 516 Flags: Print AppearanceDictionary: true Destination: 143 Annotation: Subtype: Link Rect: 381, 508, 385, 495 Flags: Print AppearanceDictionary: true Destination: 110 Thumb: true Page: Sequence: 9 Annotations: Annotation: Subtype: Link Rect: 546, 95, 552, 83 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 499, 85, 503, 73 Flags: Print AppearanceDictionary: true Destination: 204 Annotation: Subtype: Link Rect: 521, 85, 526, 73 Flags: Print AppearanceDictionary: true Destination: 219 Annotation: Subtype: Link Rect: 389, 55, 395, 43 Flags: Print AppearanceDictionary: true Destination: 319 Thumb: true Page: Sequence: 10 Annotations: Annotation: Subtype: Link Rect: 204, 221, 210, 209 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 527, 211, 533, 199 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 451, 191, 457, 179 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 225, 141, 230, 129 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 353, 91, 359, 79 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 361, 91, 367, 79 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 285, 61, 289, 49 Flags: Print AppearanceDictionary: true Destination: 219 Annotation: Subtype: Link Rect: 527, 486, 532, 474 Flags: Print AppearanceDictionary: true Destination: 204 Annotation: Subtype: Link Rect: 416, 426, 420, 414 Flags: Print AppearanceDictionary: true Destination: 143 Annotation: Subtype: Link Rect: 437, 426, 441, 414 Flags: Print AppearanceDictionary: true Destination: 219 Annotation: Subtype: Link Rect: 306, 366, 313, 356 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 306, 350, 313, 341 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 306, 339, 313, 329 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 306, 327, 309, 318 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 306, 316, 313, 306 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 306, 304, 313, 294 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 306, 292, 313, 283 Flags: Print AppearanceDictionary: true Destination: 319 Thumb: true Page: Sequence: 11 Annotations: Annotation: Subtype: Link Rect: 196, 724, 201, 712 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 342, 654, 348, 642 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 308, 534, 313, 522 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 316, 534, 322, 522 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 397, 524, 403, 512 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 213, 474, 218, 462 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 221, 464, 227, 452 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 471, 412, 477, 400 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 264, 402, 268, 390 Flags: Print AppearanceDictionary: true Destination: 219 Annotation: Subtype: Link Rect: 275, 320, 281, 308 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 493, 310, 499, 298 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 368, 300, 371, 288 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 534, 270, 540, 258 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 339, 238, 345, 226 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 263, 158, 268, 146 Flags: Print AppearanceDictionary: true Destination: 330 Thumb: true Page: Sequence: 12 Annotations: Annotation: Subtype: Link Rect: 163, 694, 168, 682 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 298, 594, 304, 582 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 306, 594, 312, 582 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 315, 594, 321, 582 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 295, 544, 299, 532 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 472, 484, 478, 472 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 480, 484, 486, 472 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 313, 374, 318, 362 Flags: Print AppearanceDictionary: true Destination: 319 Annotation: Subtype: Link Rect: 549, 54, 552, 42 Flags: Print AppearanceDictionary: true Destination: 319 Thumb: true Page: Sequence: 13 Annotations: Annotation: Subtype: Link Rect: 254, 704, 260, 692 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 243, 654, 248, 642 Flags: Print AppearanceDictionary: true Destination: 330 Annotation: Subtype: Link Rect: 364, 644, 369, 632 Flags: Print AppearanceDictionary: true Destination: 330 Thumb: true Page: Sequence: 14 Annotations: Annotation: Subtype: Link Rect: 370, 343, 474, 330 Flags: Print AppearanceDictionary: true Annotation: Subtype: Link Rect: 344, 219, 504, 207 Flags: Print AppearanceDictionary: true Thumb: true Checksum: c54ada08 Type: CRC32 Checksum: c689f5ced5a6f276a5a1583785d63407 Type: MD5 Checksum: 235f7d45afd65dc53c3adaf217f7f1ab944afb67 Type: SHA-1 Checksum: 3edcb5fcbf154ea3962d056cd9c7a663c9d012f1f5ed510b4380d1f2ca22fa1a Type: SHA-256