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The YHS-Domain of an Adenylyl Cyclase from Mycobacterium phlei Is a Probable Copper-Sensor Module.

Linder JU - PLoS ONE (2015)

Bottom Line: They are found in a variety of enzymes including copper-transporting ATPases and adenylyl cyclases.With Mg2+ as the probable physiological cofactor of the adenylyl cyclase Cu2+ specifically switches the enzyme from an inactive to an active state.By analogy to TRASH-domains binding of Cu2+ probably occurs via one conserved aspartate and three conserved cysteine-residues in the YHS-domain.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Bayreuth, Bayreuth, Germany.

ABSTRACT
YHS-domains are small protein modules which have been proposed to bind transition-metal ions like the related TRASH-domains. They are found in a variety of enzymes including copper-transporting ATPases and adenylyl cyclases. Here we investigate a class IIIc adenylyl cyclase from Mycobacterium phlei which contains a C-terminal YHS-domain linked to the catalytic domain by a peptide of 8 amino acids. We expressed the isolated catalytic domain and the full-length enzyme in E. coli. The catalytic domain requires millimolar Mn2+ as a cofactor for efficient production of cAMP, is unaffected by low micromolar concentrations of Cu2+ and inhibited by concentrations higher than 10 μM. The full-length enzyme also requires Mn2+ in the absence of an activator. However, 1-10 μM Cu2+ stimulate the M. phlei adenylyl cyclase sixfold when assayed with Mn2+. With Mg2+ as the probable physiological cofactor of the adenylyl cyclase Cu2+ specifically switches the enzyme from an inactive to an active state. Other transition-metal ions do not elicit activity with Mg2+. We favor the view that the YHS-domain of M. phlei adenylyl cyclase acts as a sensor for copper ions and signals elevated levels of the transition-metal via cAMP. By analogy to TRASH-domains binding of Cu2+ probably occurs via one conserved aspartate and three conserved cysteine-residues in the YHS-domain.

No MeSH data available.


Related in: MedlinePlus

Sequence analysis of YHS-domains in ACs.(A) Modular organisation of ACs containing YHS-domains. Amino acid numbering refers to the AC from M. phlei. (B) Alignment of YHS-, YHS/TRASH- and TRASH-domains from various bacterial proteins. The four conserved residues implicated in transition-metal binding are shaded black. Other conserved residues are shaded grey. M. phlei AC, GenBank EID14989.1; Mycobacterium marinum AC, GenBank ACC39874.1; Rhizobium leguminosarum AC, GenBank KEC71354.1; Fulvivirga imtechensis AC, GenBank ELR73472.1; Rhizobium leguminosarum Cu-ATPase, GenBank AAD26860.1; Mesorhizobium loti P-type ATPase, GenBank BAB51797.1; Pseudomonas mendocina toluene-4-hydroxylase, GenBank AAA25999.1; Halovivax ruber permease, GenBank AGB15007.1; Haloarcula marismortui ribosomal protein L24e, GenBank AAV45180.1; Desulfurococcus fermentans YHS-protein, GenBank AFL66382.1.
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pone.0141843.g001: Sequence analysis of YHS-domains in ACs.(A) Modular organisation of ACs containing YHS-domains. Amino acid numbering refers to the AC from M. phlei. (B) Alignment of YHS-, YHS/TRASH- and TRASH-domains from various bacterial proteins. The four conserved residues implicated in transition-metal binding are shaded black. Other conserved residues are shaded grey. M. phlei AC, GenBank EID14989.1; Mycobacterium marinum AC, GenBank ACC39874.1; Rhizobium leguminosarum AC, GenBank KEC71354.1; Fulvivirga imtechensis AC, GenBank ELR73472.1; Rhizobium leguminosarum Cu-ATPase, GenBank AAD26860.1; Mesorhizobium loti P-type ATPase, GenBank BAB51797.1; Pseudomonas mendocina toluene-4-hydroxylase, GenBank AAA25999.1; Halovivax ruber permease, GenBank AGB15007.1; Haloarcula marismortui ribosomal protein L24e, GenBank AAV45180.1; Desulfurococcus fermentans YHS-protein, GenBank AFL66382.1.

Mentions: In the present study we explored the function of the YHS-domain. The YHS-domain is a small cytosolic protein domain of ca. 50 amino acids named after three conserved amino acid residues, i.e. tyrosine, histidine, serine. It has first been published as part of the InterPro database (InterPro IPR007029). The YHS-domain shares extensive similarity to the TRASH domain, a protein domain binding transition-metal ions via conserved cysteine residues [14]. Due to the similarity of YHS to TRASH many domains in InterPro are annotated as both, YHS and TRASH (Fig 1B). Thus, the YHS-domain may be regarded as an expansion of the TRASH domain.


The YHS-Domain of an Adenylyl Cyclase from Mycobacterium phlei Is a Probable Copper-Sensor Module.

Linder JU - PLoS ONE (2015)

Sequence analysis of YHS-domains in ACs.(A) Modular organisation of ACs containing YHS-domains. Amino acid numbering refers to the AC from M. phlei. (B) Alignment of YHS-, YHS/TRASH- and TRASH-domains from various bacterial proteins. The four conserved residues implicated in transition-metal binding are shaded black. Other conserved residues are shaded grey. M. phlei AC, GenBank EID14989.1; Mycobacterium marinum AC, GenBank ACC39874.1; Rhizobium leguminosarum AC, GenBank KEC71354.1; Fulvivirga imtechensis AC, GenBank ELR73472.1; Rhizobium leguminosarum Cu-ATPase, GenBank AAD26860.1; Mesorhizobium loti P-type ATPase, GenBank BAB51797.1; Pseudomonas mendocina toluene-4-hydroxylase, GenBank AAA25999.1; Halovivax ruber permease, GenBank AGB15007.1; Haloarcula marismortui ribosomal protein L24e, GenBank AAV45180.1; Desulfurococcus fermentans YHS-protein, GenBank AFL66382.1.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4626032&req=5

pone.0141843.g001: Sequence analysis of YHS-domains in ACs.(A) Modular organisation of ACs containing YHS-domains. Amino acid numbering refers to the AC from M. phlei. (B) Alignment of YHS-, YHS/TRASH- and TRASH-domains from various bacterial proteins. The four conserved residues implicated in transition-metal binding are shaded black. Other conserved residues are shaded grey. M. phlei AC, GenBank EID14989.1; Mycobacterium marinum AC, GenBank ACC39874.1; Rhizobium leguminosarum AC, GenBank KEC71354.1; Fulvivirga imtechensis AC, GenBank ELR73472.1; Rhizobium leguminosarum Cu-ATPase, GenBank AAD26860.1; Mesorhizobium loti P-type ATPase, GenBank BAB51797.1; Pseudomonas mendocina toluene-4-hydroxylase, GenBank AAA25999.1; Halovivax ruber permease, GenBank AGB15007.1; Haloarcula marismortui ribosomal protein L24e, GenBank AAV45180.1; Desulfurococcus fermentans YHS-protein, GenBank AFL66382.1.
Mentions: In the present study we explored the function of the YHS-domain. The YHS-domain is a small cytosolic protein domain of ca. 50 amino acids named after three conserved amino acid residues, i.e. tyrosine, histidine, serine. It has first been published as part of the InterPro database (InterPro IPR007029). The YHS-domain shares extensive similarity to the TRASH domain, a protein domain binding transition-metal ions via conserved cysteine residues [14]. Due to the similarity of YHS to TRASH many domains in InterPro are annotated as both, YHS and TRASH (Fig 1B). Thus, the YHS-domain may be regarded as an expansion of the TRASH domain.

Bottom Line: They are found in a variety of enzymes including copper-transporting ATPases and adenylyl cyclases.With Mg2+ as the probable physiological cofactor of the adenylyl cyclase Cu2+ specifically switches the enzyme from an inactive to an active state.By analogy to TRASH-domains binding of Cu2+ probably occurs via one conserved aspartate and three conserved cysteine-residues in the YHS-domain.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Bayreuth, Bayreuth, Germany.

ABSTRACT
YHS-domains are small protein modules which have been proposed to bind transition-metal ions like the related TRASH-domains. They are found in a variety of enzymes including copper-transporting ATPases and adenylyl cyclases. Here we investigate a class IIIc adenylyl cyclase from Mycobacterium phlei which contains a C-terminal YHS-domain linked to the catalytic domain by a peptide of 8 amino acids. We expressed the isolated catalytic domain and the full-length enzyme in E. coli. The catalytic domain requires millimolar Mn2+ as a cofactor for efficient production of cAMP, is unaffected by low micromolar concentrations of Cu2+ and inhibited by concentrations higher than 10 μM. The full-length enzyme also requires Mn2+ in the absence of an activator. However, 1-10 μM Cu2+ stimulate the M. phlei adenylyl cyclase sixfold when assayed with Mn2+. With Mg2+ as the probable physiological cofactor of the adenylyl cyclase Cu2+ specifically switches the enzyme from an inactive to an active state. Other transition-metal ions do not elicit activity with Mg2+. We favor the view that the YHS-domain of M. phlei adenylyl cyclase acts as a sensor for copper ions and signals elevated levels of the transition-metal via cAMP. By analogy to TRASH-domains binding of Cu2+ probably occurs via one conserved aspartate and three conserved cysteine-residues in the YHS-domain.

No MeSH data available.


Related in: MedlinePlus