4d7k

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Revision as of 07:51, 14 September 2022

Crystal structure of N,N-8-amino-8-demethyl-D-riboflavin dimethyltransferase (RosA) from Streptomyces davawensis

Structural highlights

4d7k is a 6 chain structure with sequence from Streptomyces davaonensis JCM 4913. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

[ROSA_STRDJ] Catalyzes the S-adenosyl methionine-dependent conversion of 8-amino-8-demethyl-D-riboflavin (AF) into 8-methylamino-8-demethyl-D-riboflavin (MAF) and roseoflavin (RoF), the last two steps in the biosynthesis of the antibiotic roseoflavin.^[1] ^[2]

Publication Abstract from PubMed

N,N-8-demethyl-8-amino-D-riboflavin dimethyltransferase (RosA) catalyzes the final dimethylation of 8-demethyl-8-amino-D-riboflavin (AF) to the antibiotic roseoflavin (RoF) in Streptomyces davawensis. In the present study we have solved the X-ray structure of RosA and determined the binding properties of substrates and products. Moreover, we have used steady-state and rapid reaction kinetics to obtain detailed information on the reaction mechanism. The structure of RosA was found to be similar to previously determined S-adenosylmethionine (SAM)-dependent methyltransferases featuring two domains, a mainly alpha-helical "orthogonal bundle" and a Rossmann-like domain (alpha/beta twisted open-sheet). Bioinformatics and molecular modelling enabled us to predict the potential SAM and AF binding sites in RosA suggesting that both substrates, AF and SAM, can bind independently to their respective binding pocket. This was confirmed by kinetic experiments that demonstrated a random-order bi-bi reaction mechanism. Furthermore, we have determined the dissociation constants for substrates and products by either isothermal titration calorimetry or UV-Vis absorption spectroscopy. This revealed that both products, RoF and S-adenosylhomocysteine (SAH), bind more tightly to RosA when compared to the substrates, AF and SAM. This suggests that RosA may contribute to roseoflavin resistance in S. davawensis. The tighter binding of products is also reflected by inhibition experiments, i.e. RoF and SAH behave as competitive inhibitors for AF and SAM, respectively. We could also show that formation of a ternary complex of RosA, RoF and SAH (or SAM) leads to drastic spectral changes that are indicative of a hydrophobic environment. This article is protected by copyright. All rights reserved.

Structural and kinetic studies on RosA, the enzyme catalysing the methylation of 8-demethyl-8-amino-D-riboflavin to the antibiotic roseoflavin.,Tongsook C, Uhl MK, Jankowitsch F, Mack M, Gruber K, Macheroux P FEBS J. 2016 Feb 23. doi: 10.1111/febs.13690. PMID:26913589^[3]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Jankowitsch F, Kuhm C, Kellner R, Kalinowski J, Pelzer S, Macheroux P, Mack M. A novel N,N-8-amino-8-demethyl-D-riboflavin Dimethyltransferase (RosA) catalyzing the two terminal steps of roseoflavin biosynthesis in Streptomyces davawensis. J Biol Chem. 2011 Nov 4;286(44):38275-38285. doi: 10.1074/jbc.M111.292300. Epub, 2011 Sep 12. PMID:21911488 doi:http://dx.doi.org/10.1074/jbc.M111.292300
↑ Tongsook C, Uhl MK, Jankowitsch F, Mack M, Gruber K, Macheroux P. Structural and kinetic studies on RosA, the enzyme catalysing the methylation of 8-demethyl-8-amino-D-riboflavin to the antibiotic roseoflavin. FEBS J. 2016 Feb 23. doi: 10.1111/febs.13690. PMID:26913589 doi:http://dx.doi.org/10.1111/febs.13690
↑ Tongsook C, Uhl MK, Jankowitsch F, Mack M, Gruber K, Macheroux P. Structural and kinetic studies on RosA, the enzyme catalysing the methylation of 8-demethyl-8-amino-D-riboflavin to the antibiotic roseoflavin. FEBS J. 2016 Feb 23. doi: 10.1111/febs.13690. PMID:26913589 doi:http://dx.doi.org/10.1111/febs.13690

1 Structural highlights
2 Function
3 Publication Abstract from PubMed
4 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/4d7k"

Categories: Large Structures | Streptomyces davaonensis JCM 4913 | Gruber K | Uhl MK

@@ Line 3: / Line 3: @@
 <StructureSection load='4d7k' size='340' side='right'caption='[[4d7k]], [[Resolution|resolution]] 2.22&Aring;' scene=''>
 == Structural highlights ==
-<table><tr><td colspan='2'>[[4d7k]] is a 6 chain structure with sequence from [http://en.wikipedia.org/wiki/Strdj Strdj]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4D7K OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4D7K FirstGlance]. <br>
+<table><tr><td colspan='2'>[[4d7k]] is a 6 chain structure with sequence from [https://en.wikipedia.org/wiki/Streptomyces_davaonensis_JCM_4913 Streptomyces davaonensis JCM 4913]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4D7K OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4D7K FirstGlance]. <br>
-</td></tr><tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4d7k FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4d7k OCA], [http://pdbe.org/4d7k PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4d7k RCSB], [http://www.ebi.ac.uk/pdbsum/4d7k PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4d7k ProSAT]</span></td></tr>
+</td></tr><tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=4d7k FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4d7k OCA], [https://pdbe.org/4d7k PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4d7k RCSB], [https://www.ebi.ac.uk/pdbsum/4d7k PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4d7k ProSAT]</span></td></tr>
 </table>
+== Function ==
+[[https://www.uniprot.org/uniprot/ROSA_STRDJ ROSA_STRDJ]] Catalyzes the S-adenosyl methionine-dependent conversion of 8-amino-8-demethyl-D-riboflavin (AF) into 8-methylamino-8-demethyl-D-riboflavin (MAF) and roseoflavin (RoF), the last two steps in the biosynthesis of the antibiotic roseoflavin.<ref>PMID:21911488</ref> <ref>PMID:26913589</ref>
 <div style="background-color:#fffaf0;">
 == Publication Abstract from PubMed ==
@@ Line 20: / Line 22: @@
 </StructureSection>
 [[Category: Large Structures]]
-[[Category: Strdj]]
+[[Category: Streptomyces davaonensis JCM 4913]]
-[[Category: Gruber, K]]
+[[Category: Gruber K]]
-[[Category: Uhl, M K]]
+[[Category: Uhl MK]]
-[[Category: Alpha/beta twisted open-sheet]]
-[[Category: Flavin]]
-[[Category: Methyltransferase]]
-[[Category: Riboflavin]]
-[[Category: Roseoflavin biosynthesis]]
-[[Category: Transferase]]

4d7k

From Proteopedia

Revision as of 07:51, 14 September 2022

Crystal structure of N,N-8-amino-8-demethyl-D-riboflavin dimethyltransferase (RosA) from Streptomyces davawensis

Structural highlights

Function

Publication Abstract from PubMed

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

Views

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