7pxw
From Proteopedia
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== Function == | == Function == | ||
| - | [https://www.uniprot.org/uniprot/ | + | [https://www.uniprot.org/uniprot/LP9A_PANSI LP9A_PANSI] Lytic polysaccharide monooxygenase (LPMO) that depolymerizes crystalline and amorphous polysaccharides via the oxidation of scissile alpha- or beta-(1-4)-glycosidic bonds, yielding C1 or C4 oxidation product (PubMed:26928935, PubMed:29057953, PubMed:32818374). Catalysis by LPMOs requires the reduction of the active-site copper from Cu(II) to Cu(I) by a reducing agent and H(2)O(2) or O(2) as a cosubstrate (PubMed:26928935, PubMed:29057953). Is able to cleave phosphoric acid swollen cellulose (PASC) in the presence of a reducing agent, yielding a range of cellooligosaccharides dominated by cellobiose and cellotriose (PubMed:26928935). Activity is less sensitive to the reducing agent potential when cleaving xylan, suggesting that distinct catalytic mechanisms exist for xylan and glucan cleavage (PubMed:29057953).<ref>PMID:26928935</ref> <ref>PMID:29057953</ref> <ref>PMID:32818374</ref> |
<div style="background-color:#fffaf0;"> | <div style="background-color:#fffaf0;"> | ||
== Publication Abstract from PubMed == | == Publication Abstract from PubMed == | ||
| - | The recently discovered lytic polysaccharide monooxygenases (LPMOs) are Cu-containing enzymes capable of degrading polysaccharide substrates oxidatively. The generally accepted first step in the LPMO reaction is the reduction of the active-site metal ion from Cu(2+) to Cu(+). Here we have used a systematic diffraction data collection method to monitor structural changes in two AA9 LPMOs, one from Lentinus similis (LsAA9_A) and one from Thermoascus auranti-acus (TaAA9_A), as the active-site Cu is photoreduced in the X-ray beam. For LsAA9_A, the protein produced in two different recombinant systems was crystallized to probe the effect of post-translational modifications and different crystallization conditions on the active site and metal photoreduction. We can recommend that crystallographic studies of AA9 LPMOs wishing to address the Cu(2+) form use a total X-ray dose below 3 x 10(4) Gy, while the Cu(+) form can be attained using 1 x 10(6) Gy. In all cases, we observe the transition from a hexa-coordinated Cu site with two solvent-facing ligands to a T-shaped geometry with no exogenous ligands, and a clear increase of the | + | The recently discovered lytic polysaccharide monooxygenases (LPMOs) are Cu-containing enzymes capable of degrading polysaccharide substrates oxidatively. The generally accepted first step in the LPMO reaction is the reduction of the active-site metal ion from Cu(2+) to Cu(+). Here we have used a systematic diffraction data collection method to monitor structural changes in two AA9 LPMOs, one from Lentinus similis (LsAA9_A) and one from Thermoascus auranti-acus (TaAA9_A), as the active-site Cu is photoreduced in the X-ray beam. For LsAA9_A, the protein produced in two different recombinant systems was crystallized to probe the effect of post-translational modifications and different crystallization conditions on the active site and metal photoreduction. We can recommend that crystallographic studies of AA9 LPMOs wishing to address the Cu(2+) form use a total X-ray dose below 3 x 10(4) Gy, while the Cu(+) form can be attained using 1 x 10(6) Gy. In all cases, we observe the transition from a hexa-coordinated Cu site with two solvent-facing ligands to a T-shaped geometry with no exogenous ligands, and a clear increase of the theta(2) parameter and a decrease of the theta(3) parameter by averages of 9.2 degrees and 8.4 degrees , respectively, but also a slight increase in theta(T). Thus, the theta(2) and theta(3) parameters are helpful diagnostics for the oxidation state of the metal in a His-brace protein. On binding of cello-oligosaccharides to LsAA9_A, regardless of the production source, the theta(T) parameter increases, making the Cu site less planar, while the active-site Tyr-Cu distance decreases reproducibly for the Cu(2+) form. Thus, the theta(T) increase found on copper reduction may bring LsAA9_A closer to an oligosaccharide-bound state and contribute to the observed higher affinity of reduced LsAA9_A for cellulosic substrates. |
| - | Changes in active-site geometry on X-ray photoreduction of a lytic polysaccharide monooxygenase active-site copper and saccharide binding.,Tandrup T, Muderspach SJ, Banerjee S, Santoni G, Ipsen JO, Hernandez-Rollan C, Norholm MHH, Johansen KS, Meilleur F, Lo Leggio L IUCrJ. 2022 Aug 17;9(Pt 5):666-681. doi: 10.1107/S2052252522007175. eCollection, 2022 Sep 1. PMID:36071795<ref>PMID:36071795</ref> | + | Changes in active-site geometry on X-ray photoreduction of a lytic polysaccharide monooxygenase active-site copper and saccharide binding.,Tandrup T, Muderspach SJ, Banerjee S, Santoni G, Ipsen JO, Hernandez-Rollan C, Norholm MHH, Johansen KS, Meilleur F, Lo Leggio L IUCrJ. 2022 Aug 17;9(Pt 5):666-681. doi: 10.1107/S2052252522007175. eCollection , 2022 Sep 1. PMID:36071795<ref>PMID:36071795</ref> |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
Current revision
LPMO, expressed in E.coli, in complex with Cellotetraose
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