Photochromic Biliproteins from the Cyanobacterium <i>Anabaena </i>sp. PCC 7120:  Lyase Activities, Chromophore Exchange, and Photochromism in Phytochrome AphA

Zhao, Kai‐Hong; Ran, Yong; Li, Mei; Sun, Ye; Zhou, Ming; Storf, Max; Kupka, M.; Böhm, Stefan; Bubenzer, Claudia; Scheer, Hugo

doi:10.1021/bi0491548

Cited by 35 publications

(56 citation statements)

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“…Astonishingly, the interface between the PAS and GAF domains forms a trefoil knot ( Figure 3A) centered on the conserved Ile35, a residue that lies between Cys24 and the start of the PAS domain (Zhao et al, 2004;Wagner et al, 2005). The sequence between Cys24 and the start of the PAS domain is passed through a loop formed by a large insertion within the GAF domain .…”

Section: Structure and Assembly Of The Phytochrome Photosensory Corementioning

confidence: 99%

“…The same is true for cyanobacterial, fungal, and bacteriophytochromes (Karniol et al, 2005). While recent work implicates the importance of the P2 domain for proper holoprotein assembly of a cyanobacterial phytochrome (Zhao et al, 2004), the more distantly related phytochromes of the Cph2 subfamily lack this domain altogether but are nevertheless able to support bilin attachment (Wu and Lagarias, 2000). This suggests that the P2 domain performs an accessory role in holoprotein assembly, possibly by stabilizing CURRENT PERSPECTIVE ESSAY Figure 1.…”

mentioning

confidence: 93%

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The Structure of Phytochrome: A Picture Is Worth a Thousand Spectra

2005

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Section: Structure and Assembly Of The Phytochrome Photosensory Corementioning

confidence: 99%

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confidence: 93%

The Structure of Phytochrome: A Picture Is Worth a Thousand Spectra

2005

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“…The only biliprotein lyases that had previously been characterized mechanistically in some detail are of the E/F-type (7,17,18). In the following discussion, these two types will be compared with each other and with the phytochromes and the related cyano(bacterio)chromes that bind bilin chromophores autocatalytically (24,(45)(46)(47).…”

Section: Discussionmentioning

confidence: 99%

“…One class comprises the phytochromes and the related cyano(bacterio)chromes, which have the lyase function integrated into the apo-protein (24,(45)(46)(47)(48)(49). The other autocatalytically assembling chromoprotein is the core-membrane linker of phycobilisomes, ApcE (55).…”

Section: Discussionmentioning

confidence: 99%

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Catalytic Mechanism of S-type Phycobiliprotein Lyase

Kupka¹,

Zhang

et al. 2009

Journal of Biological Chemistry

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We now show that CpcS1 binds PCB and PEB rapidly with bi-exponential kinetics (38/119 and 12/8300 ms, respectively). Chromophore binding to the lyase is reversible and much faster than the spontaneous, but low fidelity chromophore addition to the apo-protein in the absence of the lyase. This indicates kinetic control by the enzyme, which then transfers the chromophore to the apo-protein in a slow (tens of minutes) but stereo-and regioselectively corrects the reaction. This mode of action is reminiscent of chaperones but does not require ATP. The amino acid residues Arg-18 and Arg-149 of the lyase are essential for chromophore attachment in vitro and in Escherichia coli, mutations of His-21, His-22, Trp-75, Trp-140, and Arg-147 result in reduced activity (<30% of wild type in vitro). Mutants R147Q and W69M were active but had reduced capacity for PCB binding; additionally, with W69M there was loss of fidelity in chromophore attachment. Imidazole is a noncompetitive inhibitor, supporting a bilin-binding function of histidine. Evidence was obtained that CpcS1 also catalyzes exchange of C-␤84-bound PCB in biliproteins by PEB.Phycobiliproteins are a homologous family of light-harvesting proteins present in cyanobacteria, red algae, and cryptophytes that absorb light in the spectral region between the chlorophyll absorption maxima at ϳ430 and ϳ680 nm: they contain linear tetrapyrroles (phycobilins) of which 1-4 are covalently attached to the subunits by thioether bonds to conserved cysteines (1-4). The chromophores are derived from heme by oxidative ring-opening and subsequent reduction (5) and then attached post-translationally. The correct attachment of most chromophores is catalyzed by lyases. Three phylogenetically unrelated types of lyases have been characterized in cyanobacteria (6). They are specific for certain binding sites and chromophores (7-16), but the reaction mechanisms are still unclear. A chaperone-like action has been proposed for E/Ftype lyases that catalyze chromophore attachment to Cys-84 of ␤-subunits of phyco(erythro)cyanins (17, 18). A more highly evolved function is suggested, however, by concomitant isomerizing reactions catalyzed by certain lyases (12,19,20), and by the finding that most lyases can bind the chromophore and then transfer it to the acceptor protein (15,21).Chromophore binding is particularly pronounced with the S-type lyases. CpcS1 from Nostoc PCC7120 6 rapidly forms an adduct with phycocyanobilin (PCB), 7 and the latter can be transferred in a much slower reaction to cysteine 84 of the ␤-subunits of phycocyanin (CpcB) or phycoerythrocyanin (PecB), suggesting that PCB-CpcS1 is an intermediate of the enzymatic reaction (21). Model reactions of PCB with nucleophiles resulted in the formation of addition products in which the chromophore is isomerized and which are also substrates for a CpcS1-catalzed chromophore transfer to the apo-proteins (20). We now report chromophore binding kinetics obtained by stopped-flow techniques, and identification of functional amino acid residues f...

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A rapid and robust assay for the determination of the amino acid hypusine as a possible biomarker for a high-throughput screening of antimalarials and for the diagnosis and therapy of different diseases

et al. 2011

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Eukaryotic initiation factor 5A (eIF5A) has recently been identified as a biomarker of prognostic significance and therapeutic potential for the treatment in hepatocellular carcinoma. This prompted us to establish a rapid and robust assay to determine deoxyhypusine and hypusine formed with the purified enzymes deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH) from Plasmodium to develop a rapid screening assay for antimalarial drugs. The peptide hydrolysate obtained from hypusinylated eIF5A was analyzed by ultra performance liquid chromatography (UPLC) with retention times for deoxyhypusine of 7.44 min and for hypusine of 7.30 min, respectively. The limit of detection for both compounds was 0.144 ng/μl. Determination of the specific activity of Plasmodium DOHH resulted in a twofold higher specific activity than its human counterpart. Following the iron-complexing strategy of the ferrous iron which is present in the active site of Plasmodium DOHH, a series of iron chelating compounds was tested. 2,2'-Dipyridyl and mimosine abolished DOHH activity completely while 4-oxo-piperidine-carboxylates i.e. the nitrophenylether JK8-2 and EHW 437, the oxime ether of the piperidine aldehyde, showed no inhibition although they were highly active in in vitro cultures of Plasmodium and in vivo in a rodent mouse model. The method allows a high-throughput screening (HPTS) of antimalarial drugs and the evaluation of eIF5A as a biomarker.

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Photochromic Biliproteins from the Cyanobacterium Anabaena sp. PCC 7120: Lyase Activities, Chromophore Exchange, and Photochromism in Phytochrome AphA

Cited by 35 publications

References 65 publications

The Structure of Phytochrome: A Picture Is Worth a Thousand Spectra

The Structure of Phytochrome: A Picture Is Worth a Thousand Spectra

Catalytic Mechanism of S-type Phycobiliprotein Lyase

A rapid and robust assay for the determination of the amino acid hypusine as a possible biomarker for a high-throughput screening of antimalarials and for the diagnosis and therapy of different diseases

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