Further insight into the inhibitory action of a LIM/double zinc-finger motif of an agmatinase-like protein

Cofre, Jaime; Montes, Paola; Vallejos, Alejandro; Benı́tez, José J.; García, David; Martínez‐Oyanedel, José; Carvajal, Nelson; Uribe, Elena

doi:10.1016/j.jinorgbio.2013.12.006

Cited by 9 publications

(13 citation statements)

References 22 publications

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“…2. The binding affinity of the wild-type enzyme is estimated to 18 nM (Table 1), in good agreement with a previous study [16]. Among the mutants all displayed some reduction in metal ion affinity, ranging from an~two-fold weaker binding (His127Ala) to a nearly ten-fold reduction (His206Ala; Table 1).…”

Section: Resultssupporting

confidence: 90%

“…In fact, the truncated variant exhibited a ten-fold higher k cat value and a three-fold decrease in the K m value for agmatine. A similar increase in reactivity was also observed in an ALP mutant where only one of the Zn 2+ ligands was removed (C453A mutation) [16]. It is thus likely that the LIM-domain functions as an autoinhibitory, regulatory entity for ALP.…”

Section: Introductionsupporting

confidence: 60%

“…In a previous study we demonstrated that wild-type ALP contains one Mn 2+ per subunit after purification, and the enzyme is catalytically active [16]. Adding extra Mn 2+ leads to a further increase in activity but the metal ion stoichiometry of the more active form could not be ascertained as dialysis returned the catalytic activity and metal ion stoichiometry to their initial values.…”

Section: Resultsmentioning

confidence: 94%

“…Protein interaction domains, including the LIM-domain, are recognized as key components in the regulatory machinery of the cell [15], and it has been proposed that proteins containing a LIM domain might function as biosensors that mediate communication between the cytosolic and the nuclear compartments within the cell [15]. We have recently demonstrated that ALP contains two Zn 2+ ions per monomer [14], and based on its cDNA sequence a plausible model for the ALP-LIMdomain was generated [16]. Interestingly, a deletion mutant lacking the LIM-domain was shown to be catalytically more active than the wild-type protein.…”

Section: Introductionmentioning

confidence: 99%

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Insight on the interaction of an agmatinase-like protein with Mn2+ activator ions

Quiñones

Cofre

Benı́tez

et al. 2015

Journal of Inorganic Biochemistry

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Section: Resultssupporting

confidence: 90%

Section: Introductionsupporting

confidence: 60%

Section: Resultsmentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Insight on the interaction of an agmatinase-like protein with Mn2+ activator ions

Quiñones

Cofre

Benı́tez

et al. 2015

Journal of Inorganic Biochemistry

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“…They are further divided into as many as four sub-groups, i.e. B1, B2, B3 and B4, depending upon their sequence homology and metal ion requirements for hydrolysis (3,84) .…”

Section: Metallo β Lactamasesmentioning

confidence: 99%

Kinetic, mechanistic, structural and spectroscopic investigations of Bimetallic Metallohydrolases

Selleck¹

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Cloning of two LIMCH1 isoforms: characterization of their distribution in rat brain and their agmatinase activity

García

Ordenes

Benı́tez

et al. 2015

Histochem Cell Biol

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Agmatine, a precursor for polyamine biosynthesis, is also associated with neurotransmitter, anticonvulsant, antineurotoxic and antidepressant actions in the brain. This molecule results from the decarboxylation of L-arginine by arginine decarboxylase, and it is hydrolyzed to urea and putrescine by agmatinase. Recently, we have described a new protein that also hydrolyzes agmatine, agmatinase-like protein (ALP), which was identified through immunohistochemical analysis in the hypothalamus and hippocampus of rats. However, its sequence differs greatly from all known agmatinases and does not contain the typical Mn(2+) ligands associated with the urea hydrolase family of proteins. ALP has a LIM-like domain close to its carboxyl terminus, and the removal of which results in a truncated variant with a tenfold increased k cat value and a threefold decreased K m value for agmatine. Analysis of the gene database revealed several transcripts, denominated LIMCH1 isoforms, with extreme 3' sequences identical to ALP. Limch1 gene products have been described as members of a multi-domain family of proteins with the biggest isoform containing a calponin homology (CH) domain at its N-terminus. Here, we cloned two LIMCH1 transcripts, one of 3177 bp and the other of 2709 bp (ALP contains 1569 bp) and analyzed LIMCH1 expression and distribution in rat brain using RT-PCR, Western blot and immunohistochemical analyses. LIMCH1 was detected mainly in the hypothalamic and hippocampal regions, which is similar to the distribution of ALP and agmatine in brain. In addition, we cloned and expressed both isoforms in E. coli and confirmed that they were catalytically active on agmatine with kinetic parameters similar to ALP. LIM domain-truncated variants of both isoforms moderately increased the k cat and catalytic efficiency. Thus, we propose that LIMCH1 is useful to regulate the intracellular concentrations of the neurotransmitter/neuromodulator, agmatine.

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Further insight into the inhibitory action of a LIM/double zinc-finger motif of an agmatinase-like protein

Cited by 9 publications

References 22 publications

Insight on the interaction of an agmatinase-like protein with Mn2+ activator ions

Insight on the interaction of an agmatinase-like protein with Mn2+ activator ions

Kinetic, mechanistic, structural and spectroscopic investigations of Bimetallic Metallohydrolases

Cloning of two LIMCH1 isoforms: characterization of their distribution in rat brain and their agmatinase activity

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