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6j28

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Revision as of 07:55, 23 October 2019

Crystal structure of the branched-chain polyamine synthase C9 mutein from Thermus thermophilus (Tth-BpsA C9) in complex with N4-aminopropylspermidine and 5'-methylthioadenosine

Structural highlights

6j28 is a 4 chain structure with sequence from Thet2. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Ligands:	, , ,
Gene:	bpsA, TT_C0171 (THET2)
Activity:	Transferase, with EC number 2.5.1.128
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

[Q72L89_THET2] Involved in the biosynthesis of branched-chain polyamines, which support the growth of thermophiles under high-temperature conditions. Catalyzes the sequential condensation of spermidine with the aminopropyl groups of decarboxylated S-adenosylmethionines to produce N(4)-bis(aminopropyl)spermidine via N(4)-aminopropylspermidine.[HAMAP-Rule:MF_01947]

Publication Abstract from PubMed

Branched-chain polyamine synthase (BpsA) catalyzes sequential aminopropyl transfer from the donor, decarboxylated S-adenosylmethionine (dcSAM), to the acceptor, linear-chain polyamine, resulting in the production of a quaternary-branched polyamine via tertiary branched polyamine intermediates. Here, we analyzed the catalytic properties and X-ray crystal structure of Tth-BpsA from Thermus thermophilus and compared them with those of Tk-BpsA from Thermococcus kodakarensis, which revealed differences in acceptor substrate specificity and C-terminal structure between these two enzymes. To investigate the role of the C-terminal flexible region in acceptor recognition, a region (QDEEATTY) in Tth-BpsA was replaced with that in Tk-BpsA (YDDEESSTT) to create chimeric Tth-BpsA C9, which showed a severe reduction in catalytic efficiency toward N(4) -aminopropylnorspermidine, but not toward N(4) -aminopropylspermidine, mimicking Tk-BpsA substrate specificity. Tth-BpsA C9 Tyr(346) and Thr(354) contributed to discrimination between tertiary branched-chain polyamine substrates, suggesting that the C-terminal region of BpsA recognizes acceptor substrates. Liquid chromatography-tandem mass spectrometry analysis on a Tk-BpsA reaction mixture with dcSAM revealed two aminopropyl groups bound to two of five aspartate/glutamate residues (Glu(339) , Asp(342) , Asp(343) , Glu(344) , and Glu(345) ) in the C-terminal flexible region. Mutating each of these five amino acid residues to asparagine/glutamine resulted in a slight decrease in activity. The quadruple mutant D342N/D343N/E344Q/E345Q exhibited a severe reduction in catalytic efficiency, suggesting that these aspartate/glutamate residues function to receive aminopropyl chains. In addition, the X-ray crystal structure of the Tk-BpsA ternary complex bound to N(4) -bis(aminopropyl)spermidine revealed that Asp(126) and Glu(259) interacted with the aminopropyl moiety in N(4) -aminopropylspermidine.

The C-terminal flexible region of branched-chain polyamine synthase facilitates substrate specificity and catalysis.,Hidese R, Toyoda M, Yoshino KI, Fukuda W, Wihardja GA, Kimura S, Fujita J, Niitsu M, Oshima T, Imanaka T, Mizohata E, Fujiwara S FEBS J. 2019 Jun 4. doi: 10.1111/febs.14949. PMID:31162806^[1]

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

References

↑ Hidese R, Toyoda M, Yoshino KI, Fukuda W, Wihardja GA, Kimura S, Fujita J, Niitsu M, Oshima T, Imanaka T, Mizohata E, Fujiwara S. The C-terminal flexible region of branched-chain polyamine synthase facilitates substrate specificity and catalysis. FEBS J. 2019 Jun 4. doi: 10.1111/febs.14949. PMID:31162806 doi:http://dx.doi.org/10.1111/febs.14949

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/6j28"

@@ Line 3: / Line 3: @@
 <StructureSection load='6j28' size='340' side='right'caption='[[6j28]], [[Resolution|resolution]] 1.90&Aring;' scene=''>
 == Structural highlights ==
-<table><tr><td colspan='2'>[[6j28]] is a 4 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6J28 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6J28 FirstGlance]. <br>
+<table><tr><td colspan='2'>[[6j28]] is a 4 chain structure with sequence from [http://en.wikipedia.org/wiki/Thet2 Thet2]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6J28 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6J28 FirstGlance]. <br>
 </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=FE:FE+(III)+ION'>FE</scene>, <scene name='pdbligand=MES:2-(N-MORPHOLINO)-ETHANESULFONIC+ACID'>MES</scene>, <scene name='pdbligand=MTA:5-DEOXY-5-METHYLTHIOADENOSINE'>MTA</scene>, <scene name='pdbligand=N4P:N,N-bis(3-aminopropyl)butane-1,4-diamine'>N4P</scene></td></tr>
+<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">bpsA, TT_C0171 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=262724 THET2])</td></tr>
 <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Transferase Transferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.5.1.128 2.5.1.128] </span></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=6j28 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6j28 OCA], [http://pdbe.org/6j28 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6j28 RCSB], [http://www.ebi.ac.uk/pdbsum/6j28 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6j28 ProSAT]</span></td></tr>
@@ Line 24: / Line 25: @@
 </StructureSection>
 [[Category: Large Structures]]
+[[Category: Thet2]]
 [[Category: Transferase]]
 [[Category: Fujita, J]]

6j28

From Proteopedia

Revision as of 07:55, 23 October 2019

Crystal structure of the branched-chain polyamine synthase C9 mutein from Thermus thermophilus (Tth-BpsA C9) in complex with N4-aminopropylspermidine and 5'-methylthioadenosine

Structural highlights

Function

Publication Abstract from PubMed

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

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