249 lines
29 KiB
XML
249 lines
29 KiB
XML
<document id="31598114C2A54087ADB634B3291CE40C" ID-DOI="10.1016/j.phytochem.2020.112527" ID-ISSN="1873-3700" ID-Zenodo-Dep="8292530" IM.bibliography_approvedBy="julia" IM.illustrations_approvedBy="felipe" IM.materialsCitations_approvedBy="felipe" IM.metadata_approvedBy="felipe" IM.taxonomicNames_approvedBy="julia" IM.treatments_approvedBy="julia" checkinTime="1693249326359" checkinUser="felipe" docAuthor="Oliveira, Simone T., Azevedo, Mayara I. G., Cunha, Rodrigo M. S., Silva, Christiana F. B., Muniz, Celli R., Jose, Monteiro-Júnior, E., R, Carneiro, omulo F., Nagano, Celso S., Girao ˜, Matheus S., Freitas, Cleverson D. T. & Grangeiro, Thalles B." docDate="2020" docId="743187C0FF845152FF9C1D6EFB40FAB8" docLanguage="en" docName="Phytochemistry.180.112527.pdf" docOrigin="Phytochemistry (112527) 180" docSource="http://dx.doi.org/10.1016/j.phytochem.2020.112527" docStyle="DocumentStyle:F36D69FC8B198FBE91029DF9C24697D3.5:Phytochemistry.2020-.journal_article" docStyleId="F36D69FC8B198FBE91029DF9C24697D3" docStyleName="Phytochemistry.2020-.journal_article" docStyleVersion="5" docTitle="Anacardium occidentale" docType="treatment" docVersion="3" lastPageNumber="9" masterDocId="8808FFB8FF8C515AFFF81863FFA4FFA6" masterDocTitle="Structural and functional features of a class VI chitinase from cashew (Anacardium occidentale L.) with antifungal properties" masterLastPageNumber="13" masterPageNumber="1" pageNumber="9" updateTime="1693405283461" updateUser="julia">
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<mods:title id="B7CD173B4ACCC3235E2DE29A7B530D29">Structural and functional features of a class VI chitinase from cashew (Anacardium occidentale L.) with antifungal properties</mods:title>
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<mods:namePart id="B3C0FFDEEDA3DAA8A1818A1DE570B349">Oliveira, Simone T.</mods:namePart>
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<mods:affiliation id="C4AA0A862406F572C2852292867D0A4A">Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Cear´a, Fortaleza, Cear´a, Brazil</mods:affiliation>
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<mods:namePart id="93DE3C14366B8E73310DA6C99F9FBE75">Azevedo, Mayara I. G.</mods:namePart>
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<mods:namePart id="146EEB92EA2EE3AC2BB2745F783BB044">Cunha, Rodrigo M. S.</mods:namePart>
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<mods:namePart id="4CDFDB566608A51138D6A7346F4FE927">Silva, Christiana F. B.</mods:namePart>
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<mods:namePart id="DECB1DA6B55AFB65D48B6DEF21EEA643">Muniz, Celli R.</mods:namePart>
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<mods:namePart id="77C330EAE8E46001CD9B6FE2A50DA2D6">Jose</mods:namePart>
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<mods:namePart id="5B2DC0EA787221A14B8427DE105BDFA1">Monteiro-Júnior, E.</mods:namePart>
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<mods:affiliation id="BAABE754C058AFC2C9EFA8AB638DEC19">* & Laborat´orio de Gen´etica Molecular, Departamento de Biologia, Universidade Federal do Cear´a, Fortaleza, Cear´a, Brazil</mods:affiliation>
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<mods:namePart id="D210D19BFBD85C379FE6BEE31AEF260C">Carneiro, omulo F.</mods:namePart>
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<mods:affiliation id="D1742EBC9F3F6AAF3E44A4EE9BC39307">Departamento de Engenharia de Pesca, Universidade Federal do Cear´a, Fortaleza, Cear´a, Brazil</mods:affiliation>
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<mods:namePart id="03A846E4DA10CBE6349E51024A1C4F7D">Nagano, Celso S.</mods:namePart>
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<mods:namePart id="EB46A3CBFE1596376826C995308F8E59">Girao ˜, Matheus S.</mods:namePart>
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<mods:namePart id="FCB43522B7EA4D81FEDCF024FDD3642E">Freitas, Cleverson D. T.</mods:namePart>
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<mods:namePart id="ABCB1DD18EA12B05BDDF6417880A6A25">Grangeiro, Thalles B.</mods:namePart>
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<mods:title id="A13B7AB69CF0E3AEF2F6BB16A7135EE6">Phytochemistry</mods:title>
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<mods:date id="D7269D9AD9C5AF7F31EB0D1918779A14">2020</mods:date>
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<mods:identifier id="D4086C83114690A7B2B45C6EFA90BC37" type="DOI">10.1016/j.phytochem.2020.112527</mods:identifier>
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<treatment id="743187C0FF845152FF9C1D6EFB40FAB8" LSID="urn:lsid:plazi:treatment:743187C0FF845152FF9C1D6EFB40FAB8" httpUri="http://treatment.plazi.org/id/743187C0FF845152FF9C1D6EFB40FAB8" lastPageNumber="9" pageId="8" pageNumber="9">
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<subSubSection id="B482655DFF845152FF9C1D6EFE3FFA9A" pageId="8" pageNumber="9" type="nomenclature">
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<paragraph id="FC2736D6FF845152FF9C1D6EFE3FFA9A" blockId="8.[100,754,1293,1312]" lastBlockId="8.[100,411,1321,1340]" pageId="8" pageNumber="9">
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<emphasis id="CEECEAC4FF845152FF9C1D6EFE3FFA9A" bold="true" italics="true" pageId="8" pageNumber="9">
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<heading id="A76F81BAFF845152FF9C1D6EFD56FA86" bold="true" box="[100,754,1293,1312]" centered="true" fontSize="36" level="1" pageId="8" pageNumber="9" reason="1">2.4. A molecular mechanism for the chitinolytic and antifungal properties</heading>
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<heading id="A76F81BAFF845152FF9C1D4AFE3FFA9A" box="[100,411,1321,1340]" fontSize="8" level="3" pageId="8" pageNumber="9" reason="8">
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of
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<taxonomicName id="3B984D55FF845152FF851D4AFF5FFA9A" ID-CoL="DB6H" ID-ENA="171929" authority="L." box="[125,251,1321,1340]" class="Magnoliopsida" family="Anacardiaceae" genus="Anacardium" kingdom="Plantae" order="Sapindales" pageId="8" pageNumber="9" phylum="Tracheophyta" rank="species" species="occidentale">A. occidentale</taxonomicName>
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class VI chitinase
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<paragraph id="FC2736D6FF845152FF7C1D02FDFCF979" blockId="8.[100,771,1377,1982]" pageId="8" pageNumber="9">
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To investigate a probable mechanism for the chitinolytic and antifungal properties of cashew class VI chitinase, a three-dimensional molecular model of its GH19 domain was generated by homology modeling (Fig. S21), the model was validated (Fig. S22 and Table S3) and molecular docking calculations were performed, in which a (GlcNAc)
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<subScript id="601C3493FF845152FF431D97FF60F9A4" attach="left" box="[187,196,1524,1538]" fontSize="6" pageId="8" pageNumber="9">4</subScript>
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oligosaccharide was docked in the substrate-binding cleft of AoChi (
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<figureCitation id="64A32A53FF845152FF491E6BFF48F9BA" box="[177,236,1544,1564]" captionStart="Fig" captionStartId="9.[100,130,1770,1787]" captionTargetBox="[226,1361,150,1741]" captionTargetId="figure-7@9.[225,1362,148,1743]" captionTargetPageId="9" captionText="Fig. 7. Three-dimensional molecular model of AoChi and its interaction with a chitin oligomer. (A) Cartoon representation of the three-dimensional molecular model of AoChi, which was generated by homology modeling. Side chains of putative catalytic residues are shown as sticks. Disulfide bonds are colored orange. (B) Detailed view of a chito-oligosaccharide (yellow) docked in the substrate-binding cleft of AoChi. Side chains of AoChi residues that probably interact with the docked ligand through hydrogen bonds (represented as yellow dotted lines) are shown as sticks (cyan). N and O atoms are colored blue and red, respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)" figureDoi="http://doi.org/10.5281/zenodo.8292546" httpUri="https://zenodo.org/record/8292546/files/figure.png" pageId="8" pageNumber="9">Fig. 7</figureCitation>
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). The three-dimensional model of AoChi showed the typical features of GH19 structures, with 11 α- helixes, several loop regions connecting the helical segments and 3 disulfide bonds (
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<figureCitation id="64A32A53FF845152FD571E23FD5CF9F5" box="[687,760,1600,1619]" captionStart="Fig" captionStartId="9.[100,130,1770,1787]" captionTargetBox="[226,1361,150,1741]" captionTargetId="figure-7@9.[225,1362,148,1743]" captionTargetPageId="9" captionText="Fig. 7. Three-dimensional molecular model of AoChi and its interaction with a chitin oligomer. (A) Cartoon representation of the three-dimensional molecular model of AoChi, which was generated by homology modeling. Side chains of putative catalytic residues are shown as sticks. Disulfide bonds are colored orange. (B) Detailed view of a chito-oligosaccharide (yellow) docked in the substrate-binding cleft of AoChi. Side chains of AoChi residues that probably interact with the docked ligand through hydrogen bonds (represented as yellow dotted lines) are shown as sticks (cyan). N and O atoms are colored blue and red, respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)" figureDoi="http://doi.org/10.5281/zenodo.8292546" httpUri="https://zenodo.org/record/8292546/files/figure.png" pageId="8" pageNumber="9">Fig. 7A</figureCitation>
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). The replacement of a conserved Cys residue by Ala (Ala
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<superScript id="0BED9B9EFF845152FD4E1E35FD6CF9C2" attach="left" box="[694,712,1622,1636]" fontSize="6" pageId="8" pageNumber="9">82</superScript>
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) was compensated by the emergence of a new Cys residue (Cys
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<superScript id="0BED9B9EFF845152FD7F1E11FD06F926" attach="left" box="[647,674,1650,1664]" fontSize="6" pageId="8" pageNumber="9">131</superScript>
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), and this new Cys residue was predicted to establish a disulfide bond with Cys
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<superScript id="0BED9B9EFF845152FD191EEDFD58F93A" attach="left" box="[737,764,1678,1692]" fontSize="6" pageId="8" pageNumber="9">146</superScript>
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, thus maintaining the conserved number of 3 disulfide linkages, a typical structural feature of plant GH19 chitinases (
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<figureCitation id="64A32A53FF845152FDFD1EA8FDE9F979" box="[517,589,1739,1759]" captionStart="Fig" captionStartId="9.[100,130,1770,1787]" captionTargetBox="[226,1361,150,1741]" captionTargetId="figure-7@9.[225,1362,148,1743]" captionTargetPageId="9" captionText="Fig. 7. Three-dimensional molecular model of AoChi and its interaction with a chitin oligomer. (A) Cartoon representation of the three-dimensional molecular model of AoChi, which was generated by homology modeling. Side chains of putative catalytic residues are shown as sticks. Disulfide bonds are colored orange. (B) Detailed view of a chito-oligosaccharide (yellow) docked in the substrate-binding cleft of AoChi. Side chains of AoChi residues that probably interact with the docked ligand through hydrogen bonds (represented as yellow dotted lines) are shown as sticks (cyan). N and O atoms are colored blue and red, respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)" figureDoi="http://doi.org/10.5281/zenodo.8292546" httpUri="https://zenodo.org/record/8292546/files/figure.png" pageId="8" pageNumber="9">Fig. 7A</figureCitation>
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).
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</paragraph>
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<paragraph id="FC2736D6FF845152FF7C1E84FA74FCDA" blockId="8.[100,771,1377,1982]" lastBlockId="8.[818,1488,148,1310]" pageId="8" pageNumber="9">
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Once the overall characteristics of the AoChi three-dimensional model were analyzed as well as the quality of its local and global stereo-chemical parameters were assessed, a chito-oligosaccharide with 4 units of GlcNAc was docked in the substrate-binding cleft of the validated model. The complex protein-carbohydrate was predicted to be stabilized by a network of hydrogen bonds, involving O and N atoms of the side chains of certain AoChi residues and O atoms of hydroxyl and carbonyl groups of the sugar units as well as the O atoms of the glycosidic bonds, linking the GlcNAc units of the oligosaccharide (
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<figureCitation id="64A32A53FF845152FCC218D3FCD9FF65" box="[826,893,176,195]" captionStart="Fig" captionStartId="9.[100,130,1770,1787]" captionTargetBox="[226,1361,150,1741]" captionTargetId="figure-7@9.[225,1362,148,1743]" captionTargetPageId="9" captionText="Fig. 7. Three-dimensional molecular model of AoChi and its interaction with a chitin oligomer. (A) Cartoon representation of the three-dimensional molecular model of AoChi, which was generated by homology modeling. Side chains of putative catalytic residues are shown as sticks. Disulfide bonds are colored orange. (B) Detailed view of a chito-oligosaccharide (yellow) docked in the substrate-binding cleft of AoChi. Side chains of AoChi residues that probably interact with the docked ligand through hydrogen bonds (represented as yellow dotted lines) are shown as sticks (cyan). N and O atoms are colored blue and red, respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)" figureDoi="http://doi.org/10.5281/zenodo.8292546" httpUri="https://zenodo.org/record/8292546/files/figure.png" pageId="8" pageNumber="9">Fig. 7B</figureCitation>
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). The side chain of Lys
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<superScript id="0BED9B9EFF845152FBB318C9FBC2FF1E" attach="left" box="[1099,1126,170,184]" fontSize="6" pageId="8" pageNumber="9">128</superScript>
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, which replaces the conserved Glu that acts as the proton donor in classical chitinases, was directed towards the ligand. The side chain of Glu
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<superScript id="0BED9B9EFF845152FBBD1882FBC4FF49" attach="right" box="[1093,1120,225,239]" fontSize="6" pageId="8" pageNumber="9">150</superScript>
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, the putative general base, was on the opposite side of the substrate-binding cleft. The distances between the Nζ atom of Lys
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<superScript id="0BED9B9EFF845152FC45197AFC7CFE81" attach="left" box="[957,984,281,295]" fontSize="6" pageId="8" pageNumber="9">128</superScript>
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and the atoms Oε1 and Oε2 of Glu
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were 8.0 Å and 7.9 Å. The relative positions of the side chains of Lys
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and Glu
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<superScript id="0BED9B9EFF845152FA621956FA11FEE5" attach="left" box="[1434,1461,309,323]" fontSize="6" pageId="8" pageNumber="9">150</superScript>
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as well as the average distance between their Nζ and Oε atoms agree to what is usually observed for the positions and average distances between the catalytic carboxyl groups in several structures of inverting GHs (
|
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<bibRefCitation id="98094B27FF845152FCC219C8FC59FE18" author="Mhlongo, N. N. & Skelton, A. A. & Kruger, G. & Soliman, M. E. S. & Williams, I. H." box="[826,1021,427,446]" pageId="8" pageNumber="9" pagination="1747 - 1755" refId="ref11549" refString="Mhlongo, N. N., Skelton, A. A., Kruger, G., Soliman, M. E. S., Williams, I. H., 2014. A critical survey of average distances between catalytic carboxyl groups in glycoside hydrolases. Proteins 82, 1747 - 1755. https: // doi. org / 10.1002 / prot. 24528." type="journal article" year="2014">Mhlongo et al., 2014</bibRefCitation>
|
||
). Furthermore, the distance between the Nζ atom of Lys
|
||
<superScript id="0BED9B9EFF845152FC9219A2FC21FE69" attach="left" box="[874,901,449,463]" fontSize="6" pageId="8" pageNumber="9">128</superScript>
|
||
and the most probable scissile
|
||
<emphasis id="CEECEAC4FF845152FB5519A5FB18FE7F" bold="true" box="[1197,1212,454,473]" italics="true" pageId="8" pageNumber="9">O</emphasis>
|
||
-glycosidic bond in the oligosaccharide docked in the substrate-binding groove of AoChi (approximately 4 Å), suggests that Lys
|
||
<superScript id="0BED9B9EFF845152FBB7199AFBCEFDA1" attach="left" box="[1103,1130,505,519]" fontSize="6" pageId="8" pageNumber="9">128</superScript>
|
||
could act as the proton donor during catalysis. Supporting this assumption, the theoretical p
|
||
<emphasis id="CEECEAC4FF845152FAC71A79FAE9FD8B" bold="true" box="[1343,1357,538,557]" italics="true" pageId="8" pageNumber="9">K</emphasis>
|
||
<subScript id="601C3493FF845152FAB51A41FAF1FD96" attach="left" box="[1357,1365,546,560]" fontSize="6" pageId="8" pageNumber="9">a</subScript>
|
||
value of the ζNH
|
||
<subScript id="601C3493FF845152FCA21A5EFCC7FDED" attach="left" box="[858,867,573,587]" fontSize="6" pageId="8" pageNumber="9">3</subScript>
|
||
<superScript id="0BED9B9EFF845152FCA21A53FCCDFD98" attach="left" box="[858,873,560,574]" fontSize="6" pageId="8" pageNumber="9">+</superScript>
|
||
group of Lys
|
||
<superScript id="0BED9B9EFF845152FC1D1A53FBA4FD98" attach="left" box="[997,1024,560,574]" fontSize="6" pageId="8" pageNumber="9">128</superScript>
|
||
, as predicted using the software PROPKA3, was 6.01, whereas the p
|
||
<emphasis id="CEECEAC4FF845152FC141A31FC5EFDC3" bold="true" box="[1004,1018,594,613]" italics="true" pageId="8" pageNumber="9">K</emphasis>
|
||
<subScript id="601C3493FF845152FC021A39FBA6FDCE" attach="left" box="[1018,1026,602,616]" fontSize="6" pageId="8" pageNumber="9">a</subScript>
|
||
for the δCOOH group of the general base Glu
|
||
<superScript id="0BED9B9EFF845152FA4C1A2FFA6BFDFC" attach="left" box="[1460,1487,588,602]" fontSize="6" pageId="8" pageNumber="9">150</superScript>
|
||
was calculated as 4.48 (Table S4). These numbers corroborate the enzymatic activity profile of AoChi as a function of pH values (
|
||
<figureCitation id="64A32A53FF845152FA871AE9FA60FD3B" box="[1407,1476,650,669]" captionStart="Fig" captionStartId="6.[100,130,1863,1880]" captionTargetBox="[226,1350,155,1834]" captionTargetId="figure-7@6.[225,1362,148,1836]" captionTargetPageId="6" captionText="Fig. 5. Effects of temperature (A), pH (B), metal ions (C) and chemical reagents (D) on the hydrolytic activity (A–D) and stability (A–B) of AoChi. Enzymatic assays were performed using the purified recombinant protein (200 μg/mL) and colloidal chitin as substrate, as described in the Methods section. In panels C and D, means that are significantly different (P <0.05; Bonferroni’s multiple comparisons test) when compared to control are indicated by asterisks." figureDoi="http://doi.org/10.5281/zenodo.8292542" httpUri="https://zenodo.org/record/8292542/files/figure.png" pageId="8" pageNumber="9">Fig. 5B</figureCitation>
|
||
). A rapid, exponential-like increase in AoChi activity was observed from pH 3.0 to pH 5.0. This could be due to the continuous increase in the deprotonated form of the carboxyl group of Glu
|
||
<superScript id="0BED9B9EFF845152FB131ABBFAA2FD40" attach="left" box="[1259,1286,728,742]" fontSize="6" pageId="8" pageNumber="9">150</superScript>
|
||
, acting as a base, and the concomitant deprotonation of the ζNH
|
||
<subScript id="601C3493FF845152FB3F1B62FB74FCA9" attach="left" box="[1223,1232,769,783]" fontSize="6" pageId="8" pageNumber="9">3</subScript>
|
||
<superScript id="0BED9B9EFF845152FB3F1A97FB72FCA4" attach="left" box="[1223,1238,756,770]" fontSize="6" pageId="8" pageNumber="9">+</superScript>
|
||
group of Lys
|
||
<superScript id="0BED9B9EFF845152FAAA1A97FAC9FCA4" attach="left" box="[1362,1389,756,770]" fontSize="6" pageId="8" pageNumber="9">128</superScript>
|
||
, acting as the proton donor. On the other hand, AoChi activity rapidly decreased beyond pH 6.0, reaching negligible values at pH 8.0. At pH 8.0, most (~99%) ζNH
|
||
<subScript id="601C3493FF845152FC571B37FC1CFCC4" attach="left" box="[943,952,852,866]" fontSize="6" pageId="8" pageNumber="9">3</subScript>
|
||
<superScript id="0BED9B9EFF845152FC571B2BFC1AFCF0" attach="left" box="[943,958,840,854]" fontSize="6" pageId="8" pageNumber="9">+</superScript>
|
||
groups of Lys
|
||
<superScript id="0BED9B9EFF845152FBBF1B24FBC6FCF3" attach="left" box="[1095,1122,839,853]" fontSize="6" pageId="8" pageNumber="9">128</superScript>
|
||
residues would be in a deprotonated form, and thus could no longer act as proton donors to sustain catalysis.
|
||
</paragraph>
|
||
<paragraph id="FC2736D6FF845152FCA91BE6FB40FAB8" blockId="8.[818,1488,148,1310]" pageId="8" pageNumber="9">
|
||
The CatD of AoChi has, besides Lys
|
||
<superScript id="0BED9B9EFF845152FB6D1B1CFB14FC2B" attach="left" box="[1173,1200,895,909]" fontSize="6" pageId="8" pageNumber="9">128</superScript>
|
||
, another 11 Lys residues, all of them with theoretical p
|
||
<emphasis id="CEECEAC4FF845152FBEF1BC2FB81FC12" bold="true" box="[1047,1061,929,948]" italics="true" pageId="8" pageNumber="9">K</emphasis>
|
||
<subScript id="601C3493FF845152FBDD1BCBFB89FC10" attach="left" box="[1061,1069,936,950]" fontSize="6" pageId="8" pageNumber="9">a</subScript>
|
||
values of their ζNH
|
||
<subScript id="601C3493FF845152FB0C1BCBFB59FC10" attach="left" box="[1268,1277,936,950]" fontSize="6" pageId="8" pageNumber="9">3</subScript>
|
||
<superScript id="0BED9B9EFF845152FB0C1BF8FAA7FC0F" attach="left" box="[1268,1283,923,937]" fontSize="6" pageId="8" pageNumber="9">+</superScript>
|
||
groups ranging from 10.18 (Lys
|
||
<superScript id="0BED9B9EFF845152FC6B1BD4FC01FC63" attach="left" box="[915,933,951,965]" fontSize="6" pageId="8" pageNumber="9">76</superScript>
|
||
) to 10.75 (Lys
|
||
<superScript id="0BED9B9EFF845152FBD31BD4FBE2FC63" attach="left" box="[1067,1094,951,965]" fontSize="6" pageId="8" pageNumber="9">111</superScript>
|
||
). The average experimental p
|
||
<emphasis id="CEECEAC4FF845152FAA21BDEFACCFC76" bold="true" box="[1370,1384,957,976]" italics="true" pageId="8" pageNumber="9">K</emphasis>
|
||
<subScript id="601C3493FF845152FA901BA7FAD4FC74" attach="left" box="[1384,1392,964,978]" fontSize="6" pageId="8" pageNumber="9">a</subScript>
|
||
values for Lys residues, measured in 157 proteins, have been determined to be 10.68 (
|
||
<bibRefCitation id="98094B27FF845152FC801B97FB89FBAE" author="Pahari, S. & Sun, L. & Alexov, E." box="[888,1069,1012,1032]" pageId="8" pageNumber="9" refId="ref12047" refString="Pahari, S., Sun, L., Alexov, E., 2019. PKAD: a database of experimentally measured pKa values of ionizable groups in proteins. Database 2019, baz 024. https: // doi. org / 10.1093 / database / baz 024." type="journal volume" year="2019">Pahari et al., 2019</bibRefCitation>
|
||
), which is close to the intrinsic p
|
||
<emphasis id="CEECEAC4FF845152FA801B96FA22FBAE" bold="true" box="[1400,1414,1013,1032]" italics="true" pageId="8" pageNumber="9">K</emphasis>
|
||
<subScript id="601C3493FF845152FA7E1B9FFA2AFBAC" attach="left" box="[1414,1422,1020,1034]" fontSize="6" pageId="8" pageNumber="9">a</subScript>
|
||
of the ζNH
|
||
<subScript id="601C3493FF845152FCA21C7BFCC7FB80" attach="left" box="[858,867,1048,1062]" fontSize="6" pageId="8" pageNumber="9">3</subScript>
|
||
<superScript id="0BED9B9EFF845152FCA21C68FCCDFBBF" attach="left" box="[858,873,1035,1049]" fontSize="6" pageId="8" pageNumber="9">+</superScript>
|
||
group of Lys in bulk water, which is 10.4 (
|
||
<bibRefCitation id="98094B27FF845152FAF41C73FCC6FB99" author="Nozaki, Y. & Tanford, C." pageId="8" pageNumber="9" pagination="736 - 742" refId="ref11988" refString="Nozaki, Y., Tanford, C., 1967. Acid-base titrations in concentrated guanidine hydrochloride. Dissociation constants of the guamidinium ion and of some amino acids. J. Am. Chem. Soc. 89, 736 - 742. https: // doi. org / 10.1021 / ja 00980 a 002." type="journal article" year="1967">Nozaki and Tanford, 1967</bibRefCitation>
|
||
). However, Lys and other ionizable residues buried in the hydrophobic environments of certain proteins might have anomalous p
|
||
<emphasis id="CEECEAC4FF845152FA411C2BFA63FBFD" bold="true" box="[1465,1479,1096,1115]" italics="true" pageId="8" pageNumber="9">K</emphasis>
|
||
<subScript id="601C3493FF845152FA3F1C33FA6BFBF8" attach="left" box="[1479,1487,1104,1118]" fontSize="6" pageId="8" pageNumber="9">a</subScript>
|
||
values, with large deviations from the intrinsic p
|
||
<emphasis id="CEECEAC4FF845152FAEB1C07FA85FBD1" bold="true" box="[1299,1313,1124,1143]" italics="true" pageId="8" pageNumber="9">K</emphasis>
|
||
<subScript id="601C3493FF845152FAD91C0FFA8DFBDC" attach="left" box="[1313,1321,1132,1146]" fontSize="6" pageId="8" pageNumber="9">a</subScript>
|
||
values in water. Indeed, buried Lys residues with experimental p
|
||
<emphasis id="CEECEAC4FF845152FB0F1CE3FAA1FB35" bold="true" box="[1271,1285,1152,1171]" italics="true" pageId="8" pageNumber="9">K</emphasis>
|
||
<subScript id="601C3493FF845152FAFD1CE4FAA9FB33" attach="left" box="[1285,1293,1159,1173]" fontSize="6" pageId="8" pageNumber="9">a</subScript>
|
||
values as low as 6.2 have been experimentally determined (
|
||
<bibRefCitation id="98094B27FF845152FB3C1CFFFA65FB09" author="Kougentakis, C. M. & Grasso, E. M. & Robinson, A. C. & Caro, J. A. & Schlessman, J. L. & Majumdar, A. & Garcia-Moreno, E. B." box="[1220,1473,1180,1199]" pageId="8" pageNumber="9" pagination="383 - 387" refId="ref10528" refString="Kougentakis, C. M., Grasso, E. M., Robinson, A. C., Caro, J. A., Schlessman, J. L., Majumdar, A., Garcia-Moreno, E. B., 2018. Anomalous properties of Lys residues buried in the hydrophobic interior of a protein revealed with 15 N-detect NMR spectroscopy. J. Phys. Chem. Lett. 9, 383 - 387. https: // doi. org / 10.1021 / acs. jpclett. 7 b 02668." type="journal article" year="2018">Kougentakis et al., 2018</bibRefCitation>
|
||
). Therefore, this analysis supports the assumption that Lys
|
||
<superScript id="0BED9B9EFF845152FAAF1CD1FAD6FB66" attach="left" box="[1367,1394,1202,1216]" fontSize="6" pageId="8" pageNumber="9">128</superScript>
|
||
of AoChi can act as the proton donor during catalysis, explaining the ability of
|
||
<taxonomicName id="3B984D55FF845152FA431CB7FC31FAA4" class="Magnoliopsida" family="Anacardiaceae" genus="Anacardium" kingdom="Plantae" order="Sapindales" pageId="8" pageNumber="9" phylum="Tracheophyta" rank="species" species="occidentale">
|
||
<emphasis id="CEECEAC4FF845152FA431CB7FA6DFB41" bold="true" box="[1467,1481,1236,1255]" italics="true" pageId="8" pageNumber="9">A</emphasis>
|
||
.
|
||
<emphasis id="CEECEAC4FF845152FCCA1C8CFC31FAA4" bold="true" box="[818,917,1263,1282]" italics="true" pageId="8" pageNumber="9">occidentale</emphasis>
|
||
</taxonomicName>
|
||
class VI chitinase to degrade colloidal chitin and to cause damages in the cell walls of pathogenic fungi.
|
||
</paragraph>
|
||
</subSubSection>
|
||
</treatment>
|
||
</document> |