The zinc-containing high Km cyclic nucleotide phosphodiesterase of bakers' yeast.

Londesborough, John; Suoranta, K

doi:10.1016/s0021-9258(18)32815-1

Cited by 33 publications

(15 citation statements)

References 26 publications

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“…The specificity constant ( k cat / K M cAMP )/( k cat / K M cGMP ) of 1.4 suggests a dual activity of yPDE1 on hydrolysis of both cAMP and cGMP, with a slightly better efficacy on cAMP than cGMP. Our K M of yPDE1 for cAMP is comparable with the early report of K M values of 100–120 μM, but our yPDE1 enzyme is much more active, as shown by a V max at least 20-fold higher than those of the previously reported proteins. ,, …”

Section: Resultssupporting

confidence: 90%

“…The native bakers’ yeast yPDE1 was shown to contain two zinc ions per molecule by an atomic absorption spectrometer . In a plain assay buffer without divalent metals, our recombinant yPDE1 shows similar enzymatic efficacy for hydrolysis of cAMP and cGMP: K M of 110 μM and k cat of 16.9 s –1 for cAMP and K M of 105 μM and k cat of 11.8 s –1 for cGMP (Table ).…”

Section: Resultsmentioning

confidence: 95%

“…The signaling of cAMP in yeast has been shown to play critical roles in physiological processes, including metabolism, cell wall biosynthesis, cell growth, and mating. − Overexpression of the PDE2 gene enhances the tolerance of yeast to oxidative and ethanol stress for survival. − Both yPDE1 and yPDE2 appear to participate in the cAMP signaling pathway, although yPDE2 shows high affinity with a K M of 0.17–1.0 μM for cAMP, − and yPDE1 has a low affinity with a K M of 100–150 μM. ,, In comparison with numerous publications about the roles of cAMP in the physiological processes of yeast, the cGMP signaling pathway has not been well studied, although it was reported that cGMP activated cAMP-dependent protein kinase of Pichia pastoris yeast . There is only one report showing that yPDE1 from Candida albicans has very weak cGMP activity with a K M of 250 μM and a V max of 0.044 μmol mg –1 min –1 , in comparison with a K M of 490 μM and a V max of 1.17 μmol mg –1 min –1 for cAMP .…”

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confidence: 99%

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Dual Specificity and Novel Structural Folding of Yeast Phosphodiesterase-1 for Hydrolysis of Second Messengers Cyclic Adenosine and Guanosine 3′,5′-Monophosphate

et al. 2014

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Cyclic nucleotide phosphodiesterases (PDEs) decompose second messengers cAMP and cGMP that play critical roles in many physiological processes. PDE1 of Saccharomyces cerevisiae has been subcloned and expressed in Escherichia coli. Recombinant yPDE1 has a KM of 110 μM and a kcat of 16.9 s–1 for cAMP and a KM of 105 μM and a kcat of 11.8 s–1 for cGMP. Thus, the specificity constant (kcat/KMcAMP)/(kcat/KMcGMP) of 1.4 indicates a dual specificity of yPDE1 for hydrolysis of both cAMP and cGMP. The crystal structures of unliganded yPDE1 and its complex with GMP at 1.31 Å resolution reveal a new structural folding that is different from those of human PDEs but is partially similar to that of some other metalloenzymes such as metallo-β-lactamase. In spite of their different structures and divalent metals, yPDE1 and human PDEs may share a common mechanism for hydrolysis of cAMP and cGMP.

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Section: Resultssupporting

confidence: 90%

Section: Resultsmentioning

confidence: 95%

mentioning

confidence: 99%

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Dual Specificity and Novel Structural Folding of Yeast Phosphodiesterase-1 for Hydrolysis of Second Messengers Cyclic Adenosine and Guanosine 3′,5′-Monophosphate

et al. 2014

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“…This is also consistent with the early reports that bakery yeast PDE2 contained zinc ion and that magnesium ion yielded the highest hydrolytic activity. 18,19 The zinc ion has an octahedral conformation and forms four coordinations with protein residues His278, His349, Asp350, and Asp462 and two waters (W1 and W2 in Figure 3). The magnesium ion also has an octahedral conformation and chelates with Asp350 and five water molecules.…”

Section: Monomeric Capde2mentioning

confidence: 99%

“…Both yPDE1 and yPDE2 hydrolyze cAMP and participate in cAMP signaling pathways. Yeast PDE2 was named as a high-affinity cAMP PDE because its K M values were in the range of 0.17–1.0 μM, − whereas yPDE1 has a low affinity with a K M value of 100–150 μM for cAMP. − In contrast with the well-characterized roles of cAMP in the physiological processes of yeast, the cGMP signaling pathway of yeasts has rarely been reported. C. albicans PDE1 was reported to have K M values of 250 and 490 μM and V max values of 0.044 and 1.17 μmol mg –1 min –1 for cGMP and cAMP, respectively, suggesting its inefficiency in the cGMP signaling pathway due to its extremely low V max .…”

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confidence: 99%

Crystal Structures of Candida albicans Phosphodiesterase 2 and Implications for Its Biological Functions

et al. 2018

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The cAMP signaling system plays important roles in the physiological processes of pathogen yeast Candida albicans, but its functional mechanism has not been well illustrated. Here, we report the enzymatic characterization and crystal structures of C. albicans phosphodiesterase 2 (caPDE2) in the unliganded and 3-isobutyl-1-methylxanthine-complexed forms. caPDE2 is a monomer in liquid and crystal states and specifically hydrolyzes cAMP with a K of 35 nM. It does not effectively hydrolyze cGMP as shown by the 1.32 × 10-fold specificity of cAMP/cGMP. The crystal structure of caPDE2 shows significant differences from those of human PDEs. First, the N-terminal fragment of caPDE2 (residues 1-201) tightly associates with the catalytic domain to form a rigid molecular entity, implying its stable molecular conformation for C. albicans to resist environmental stresses. Second, the M-loop, a critical fragment for binding of the substrate and inhibitors to human PDEs, is not a part of the caPDE2 active site. This feature of caPDE2 may provide a structural basis for the design of selective inhibitors for the treatment of yeast infection.

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Sperm motility in the horseshoe crab. V: Zinc removal mediates chelator initiation of motility

et al. 1985

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Limulus spermatozoa are nonmotile after dilution into sea water, and motility is initiated in situ by a sperm motility-initiating peptide (SMI) that emanates from Limulus eggs. In the absence of SMI, several structurally different chelators (EDTA, EGTA, phenanthroline, dipyridyl, cysteine, and dithiothreitol; 1 mM each) initiated sperm motility; and in the presence of SMI, EDTA prolonged the duration of motility initiated by SMI. Of the elements determined to be present in Limulus sperm, Zn+2 (0.1 pM free Znf2) was both the most effective at preventing EDTA initiation of motility and the only ion to prevent EDTA prolongation of SMI-initiated motility. 65Zn was concentrated from sea water by Limulus sperm and was rapidly removed by EDTA. These results indicate that removal of the Zn+2 normally associated with Limulus spermatozoa can initiate or enhance motility. However, SMI does not act as a Zn+2 chelator, since SMI removed little 65Zn from the sperm while at the same time initiating motility of greater percentage and duration than did EDTA. The mechanism by which Znf2 removal enhances sperm motility is not yet known but could involve a Znf2-dependent or Znf2-regulated step in the pathway of SMI action.In Limulus spermatozoa, motility is initiated in situ by a sperm motility-initiating peptide (SMD that emanates from eggs (Clapper and Brown, 'Boa; Clapper and Epel, '82a). This motility initiation by SMI is not mediated by the influx of any specific extracellular ion or by changes in intracellular pH (Clapper and Epel, '82b) as is the case with sea urchin sperm motility (Christen et al., '82; Lee et al., '83). However, an increase in CAMP has been correlated with motility initiation by crude SMI extracts (Tubb et a1 '79) and mobilization of intracellular Caf3 may also be involved in SMI-initiated motility (Clapper and Epel, '82b).A few previous studies have provided evidence for a possible heavy metal involvement in Limulus sperm motility. Mowbray and Brown ('74) reported that both cysteine (-10 mM) and EDTA (-1 mM) facilitated sperm motility near egg sections. However, a n apparently contradictory finding was reported jby Bishop and Nidek ('771, who found that C U +~ initiated motility in Limulus sperm. A subsequent study found that C U +~, Ni+2, Ca+2, and Mgf2 would each initiate motility, but only at levels far above those found either in sea water or in sea water containing concentrated egg extracts (Clapper and Brown, 'Bob).These findings (that motility is initiated both by metal chelators and by some metal ions) could result from the removal or displacement of a metal ion that is inhibitory to sperm motility. Indeed, the present study presents evidence that removal of Znf2 from Limulus sperm mediates the initiation of motility by metal chelators. However, experiments with 65Zn show that SMI does not act as a Zn+' chelator; therefore, chelators and SMI must have somewhat different mechanisms of action. Possibly SMI action is mediated by a Znf2-dependent or Zn+2-regulated event, and the removal of Znf...

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The zinc-containing high Km cyclic nucleotide phosphodiesterase of bakers' yeast.

Cited by 33 publications

References 26 publications

Dual Specificity and Novel Structural Folding of Yeast Phosphodiesterase-1 for Hydrolysis of Second Messengers Cyclic Adenosine and Guanosine 3′,5′-Monophosphate

Dual Specificity and Novel Structural Folding of Yeast Phosphodiesterase-1 for Hydrolysis of Second Messengers Cyclic Adenosine and Guanosine 3′,5′-Monophosphate

Crystal Structures of Candida albicans Phosphodiesterase 2 and Implications for Its Biological Functions

Sperm motility in the horseshoe crab. V: Zinc removal mediates chelator initiation of motility

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