<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>0120-2804</journal-id>
<journal-title><![CDATA[Revista Colombiana de Química]]></journal-title>
<abbrev-journal-title><![CDATA[Rev.Colomb.Quim.]]></abbrev-journal-title>
<issn>0120-2804</issn>
<publisher>
<publisher-name><![CDATA[Departamento de Química,  Universidad Nacional de Colombia.]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0120-28042019000100016</article-id>
<article-id pub-id-type="doi">10.15446/rev.colomb.quim.v48n1.75273</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Functional identification and subcellular localization of NAD kinase in the protozoan parasite Giardia intestinalis]]></article-title>
<article-title xml:lang="es"><![CDATA[Identificación funcional y localización subcelular de la NAD quinasa en el parásito protozoario Giardia intestinalis]]></article-title>
<article-title xml:lang="pt"><![CDATA[Identificação funcional e localização subcelular da NAD quinase no parasita protozoário Giardia intestinalis]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Jutinico Shubach]]></surname>
<given-names><![CDATA[Laura Marcela]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Contreras Rodríguez]]></surname>
<given-names><![CDATA[Luis Ernesto]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[García Castañeda]]></surname>
<given-names><![CDATA[Javier Eduardo]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Ramírez Hernández]]></surname>
<given-names><![CDATA[María Helena]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad Nacional de Colombia Departamento de Química ]]></institution>
<addr-line><![CDATA[Bogotá ]]></addr-line>
<country>Colombia</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Universidad Nacional de Colombia Departamento de Farmacia ]]></institution>
<addr-line><![CDATA[Bogotá ]]></addr-line>
<country>Colombia</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,Universidad Nacional de Colombia Departamento de Biología ]]></institution>
<addr-line><![CDATA[Bogotá ]]></addr-line>
<country>Colombia</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>06</month>
<year>2019</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>06</month>
<year>2019</year>
</pub-date>
<volume>48</volume>
<numero>1</numero>
<fpage>16</fpage>
<lpage>25</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_arttext&amp;pid=S0120-28042019000100016&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_abstract&amp;pid=S0120-28042019000100016&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_pdf&amp;pid=S0120-28042019000100016&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract Nicotinamide adenine dinucleotide phosphate (NADP) is an essential biomolecule that participates in the redox homeostasis and synthesis of signaling compounds. NAD kinase (NADK) (EC 2.7.1.23 / 2.7.1.86) is the only enzyme capable of synthesizing NADP. This study offers an approach to the NADP metabolism in the parasite Giardia intestinalis, the etiological agent of giardiasis, a disease of high prevalence in America, Asia and Africa. Through bioinformatics tools a NADK enzyme candidate was identified, whose tertiary structure modeling demonstrated distinctive and universal motifs of characterized NADKs. The corresponding recombinant protein (His-GINADK) was expressed in Escherichia coli BL21 (DE3) and its partial purification was achieved by nickel affinity chromatography. Functional identification, which showed NADP synthesis, was completed through enzymatic assays evaluated by RP-HPLC. A cytosolic localization of the endogenous GINADK enzyme was observed in trophozoites throughout indirect immunofluorescence analysis, using polyclonal antibodies produced in mice by its immunization with the His-GINADK protein, purified from inclusion bodies. Taken together, our results contribute to the understanding of the NADP metabolism and the physiological role of NADK in the Giardia model.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen El dinucleótido de adenina y nicotinamida fosfato (NADP) es una biomolécula esencial que participa en la homeostasis redox y en la síntesis de compuestos de señalización. La única enzima capaz de sintetizar NADP es la NAD Quinasa (NADK, EC 2.7.1.23 / 2.7.1.86). En este estudio se presenta un acercamiento al metabolismo del NADP en el parásito Giardia intestinalis, agente etiológico de la giardiasis, una enfermedad de alta prevalencia en América, África y Asia. Mediante herramientas bioinformáticas se identificó un candidato a NADK, cuya predicción a nivel de estructura terciaria mostró motivos característicos y universales de NADKs previamente caracterizadas. La proteína recombinante correspondiente (His-GINADK) se expresó en Escherichia coli BL21 (DE3) y se purificó parcialmente mediante cromatografía de afinidad a níquel. La síntesis de NADP por parte de la proteína His-GINADK se comprobó mediante ensayos enzimáticos evaluados por RP-HPLC. Adicionalmente, se determinó una localización subcelular citosólica en trofozoítos del parásito, empleando inmunofluorescencia indirecta y anticuerpos policlonales producidos en modelos murinos inmunizados con la proteína His-GINADK purificada a partir de cuerpos de inclusión. Los resultados obtenidos representan un avance en el entendimiento del metabolismo del NADP y de la importancia fisiológica de la NADK en el modelo de Giardia.]]></p></abstract>
<abstract abstract-type="short" xml:lang="pt"><p><![CDATA[Resumo A nicotinamida adenina dinucleótido fosfato (NADP) é uma biomolécula essencial que participa na homeostase redox e na síntese de importantes compostos de sinalização. A NAD quinase (NADK) (EC 2.7.1.23 / 2.7.1.86) é a única enzima capaz de sintetizar o NADP. Este estudo apresenta uma abordagem do metabolismo do NADP no parasita Giardia intestinalis que causa giardíase, uma doença de alta prevalência na América, Ásia e África. Através de ferramentas de bioinformática, um candidato a enzima NADK foi identificado no parasita, cuja modelagem de estrutura terciária, demonstra motivos distintos e universais de NADKs caracterizadas. A correspondente proteína recombinante (His-GINADK) foi expressa em Escherichia coli BL21 (DE3) e a sua purificação parcial foi conseguida por cromatografia de afinidade com níquel. A identificação funcional, que mostrou a síntese de NADP, foi completada através de ensaios enzimáticos avaliados por RP-HPLC. Uma localização citosólica da enzima GINADK endógena foi observada em trofozoítos ao longo da análise de imunofluorescência indireta, utilizando anticorpos policlonais produzidos em camundongos, imunizados com a proteína His-GINADK purificada de corpos de inclusão. Em conjunto, nossos resultados contribuem para a compreensão do metabolismo do NADP e da importância fisiológica do NADK no modelo de Giardia.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[cytosolic localization]]></kwd>
<kwd lng="en"><![CDATA[NADP synthesis]]></kwd>
<kwd lng="en"><![CDATA[recombinant protein]]></kwd>
<kwd lng="es"><![CDATA[localización citosólica]]></kwd>
<kwd lng="es"><![CDATA[síntesis de NADP]]></kwd>
<kwd lng="es"><![CDATA[proteína recombinante]]></kwd>
<kwd lng="pt"><![CDATA[localização citosólica]]></kwd>
<kwd lng="pt"><![CDATA[síntese de NADP]]></kwd>
<kwd lng="pt"><![CDATA[proteína recombinante]]></kwd>
</kwd-group>
</article-meta>
</front><back>
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