Leishmania major LmACR2 Is a Pentavalent Antimony Reductase That Confers Sensitivity to the Drug Pentostam

Arsenicals and antimonials are first line drugs for the treatment of trypanosomal and leishmanial diseases. To create the active form of the drug, Sb(V) must be reduced to Sb(III). Because arsenic and antimony are related metalloids, and arsenical resistant Leishmania strains are frequently cross-resistant to antimonials, we considered the possibility that Sb(V) is reduced by a leishmanial As(V) reductase. The sequence for the arsenate reductase of Saccharomyces cerevisiae, ScAcr2p, was used to clone the gene for a homologue, LmACR2, from Leishmania major. LmACR2 was able to complement the arsenate-sensitive phenotype of an arsC deletion strain of Escherichia coli or an ScACR2 deletion strain of Saccharomyces cerevisiae. Transfection of Leishmania infantum with LmACR2 augmented Pentostam sensitivity in intracellular amastigotes. LmACR2 was purified and shown to reduce both As(V) and Sb(V). This is the first report of an enzyme that confers Pentostam sensitivity in intracellular amastigotes of Leishmania. We propose that LmACR2 is responsible for reduction of the pentavalent antimony in Pentostam to the active trivalent form of the drug in Leishmania. Arsenicals and antimonials are first line drugs for the treatment of trypanosomal and leishmanial diseases. To create the active form of the drug, Sb(V) must be reduced to Sb(III). Because arsenic and antimony are related metalloids, and arsenical resistant Leishmania strains are frequently cross-resistant to antimonials, we considered the possibility that Sb(V) is reduced by a leishmanial As(V) reductase. The sequence for the arsenate reductase of Saccharomyces cerevisiae, ScAcr2p, was used to clone the gene for a homologue, LmACR2, from Leishmania major. LmACR2 was able to complement the arsenate-sensitive phenotype of an arsC deletion strain of Escherichia coli or an ScACR2 deletion strain of Saccharomyces cerevisiae. Transfection of Leishmania infantum with LmACR2 augmented Pentostam sensitivity in intracellular amastigotes. LmACR2 was purified and shown to reduce both As(V) and Sb(V). This is the first report of an enzyme that confers Pentostam sensitivity in intracellular amastigotes of Leishmania. We propose that LmACR2 is responsible for reduction of the pentavalent antimony in Pentostam to the active trivalent form of the drug in Leishmania. Leishmania species are distributed worldwide. During its life cycle, the parasite goes through two developmental stages. The promastigote form of the parasite resides in the intestinal tract of the insect vector. The amastigote form of the parasite resides in macrophages and other mononuclear phagocytes in the mammalian host. Between 10 and 15 million people have clinical symptoms of leishmaniasis, and 400,000 new cases are diagnosed each year. Increases in personal and business travel and intervention in regional conflicts such as the last two Iraqi wars have resulted in a significant rise in cases of Leishmania in the United States (1Lockwood D. Nat. Med. 2004; 10: 110Crossref PubMed Scopus (4) pentavalent drugs Pentostam and are the first line treatment for leishmaniasis, and to drugs is a clinical Med. PubMed Scopus PubMed Scopus To create the active form of the drug, Sb(V) is first reduced to D. PubMed in the are to Pentostam are D. PubMed D. PubMed Scopus and antimony have and are by the 2004; PubMed Scopus PubMed Scopus 2004; PubMed Scopus PubMed Scopus and PubMed Scopus D. PubMed We propose that a of the Sb(V) in Pentostam is reduced by a leishmanial As(V) reductase. are in and and are for to arsenate PubMed Scopus arsenate reductase a Leishmania enzyme related to shown to reduction of arsenate and in 2004; PubMed Scopus is a for a are that a of enzyme and drug sensitivity in Leishmania the arsenate reductase from Saccharomyces arsenate to both in and in PubMed PubMed Scopus PubMed Scopus PubMed Scopus is to the PubMed Scopus and to a PubMed Scopus and as PubMed Scopus Because of its to is to by of We used the sequence to a related Leishmania This LmACR2, was both and Escherichia coli LmACR2 the arsenate-sensitive of the ScACR2 deletion of and a deletion of the coli arsC a arsenate reductase. LmACR2 was purified and The Leishmania enzyme and for arsenate reductase with a of for As(V) and is by and LmACR2 Sb(V) to with a of To a LmACR2 and Pentostam LmACR2 was Leishmania infantum amastigotes from sensitivity to This is the first of an enzyme that is in and for of the drug Pentostam in intracellular Leishmania from and in the form of and from and of Leishmania was for an of The to ScACR2 is in and in The LmACR2, was by by as and as and the and of the for the LmACR2 was by as and as Leishmania LmACR2 was by as and as was first and to the of the in a from The rise to for in for in and for in from coli in a PubMed Scopus with as The arsenate phenotype of was in coli strains or as PubMed Scopus in of arsenate and was from the 15 of and PubMed or in a PubMed or with each of as of of of with The for infantum strain the gene D. PubMed Scopus was used for in in PubMed with 10 as and by of The promastigote used to the line as D. PubMed Scopus with and with infantum a of for and macrophages with of of and was as a D. PubMed Scopus are as of LmACR2 and coli used and purified as PubMed Scopus PubMed Scopus LmACR2 was purified from of coli strain in of with an of was to a of as and the was for an The with in 10 and a of of of and by a through a of was to the The was for and the was a of a with The was with of by with of with the of to LmACR2 was by purified LmACR2 and The from the was to a to with with 10 and with and purified from the by the by the of and PubMed Scopus and LmACR2, of reductase was by a PubMed Scopus The reductase and of was used to that the of the was in the of and as was and as of of LmACR2 by a of for The by was with two of purified of LmACR2 was with the of for the with the used for arsenate as The was with of an of and with was used for of Sb(V) and with as the a of PubMed Scopus Sb(V) Sb(V) a with The of antimony in each was by Sb(V) and of the other The for the of in the of LmACR2, was a of the with of the Sb(V) was the During the of and the used in the reduced and of Leishmania sequence for an that is in was with and as to a that was and rise to a gene that we LmACR2 LmACR2 and are in and LmACR2 with a of with ScAcr2p, and a of an active the sequence PubMed Scopus and LmACR2 the The two sequence and of LmACR2 the of an of was the in with a from or with the of the arsenate in the coli arsC deletion strain PubMed Scopus LmACR2 of was with gene in the of LmACR2 the As(V) sensitivity of an strain of coli sensitivity to Sb(V) confers arsenate in in coli was as of in LmACR2 is a that was purified by The of purified LmACR2 was by a the sequence of the LmACR2 gene with the and the gene a a of was with a as a PubMed Scopus and of LmACR2 was purified as The of LmACR2 was from its The of the are for and of each of by a of of coli strain with of of and of from an of LmACR2 to reduce arsenate to was by a for and PubMed Scopus PubMed Scopus of arsenate was by LmACR2 in the of the coli for PubMed Scopus and the coli as for LmACR2 the for arsenate was with 10 with The was with both LmACR2 in to the ScAcr2p, and an of PubMed Scopus The of trivalent arsenate reductase was with an of The arsenical and with of and and LmACR2 arsenate reductase in reductase of purified The arsenate reductase of LmACR2 was with and reductase was from the of as purified coli LmACR2 arsenate reductase of purified The arsenate reductase of LmACR2 was with and coli or The of arsenate reductase was from the of each of as The the from two of arsenate reductase by trivalent reductase and of arsenate was in the of of The and of the from the of the of the an the drugs for treatment of such as Pentostam are pentavalent antimonials that must be by reduction to the of LmACR2 to reduce Sb(V) was Pentostam is the of and We used an of trivalent and pentavalent antimonials in the of each species is by PubMed Scopus LmACR2 reduced Sb(V) to in a reductase both and The of Sb(V) reduction a of for Sb(V) and a of the for Sb(V) is for and the is that LmACR2 is a reductase with for Sb(V) Sb(V) to Sb(III). The Sb(V) reductase of LmACR2 was with purified coli and with Sb(V) and by and the of was by as reduction of Sb(V) was for the with of Sb(V) reduction from the of with The line is the to the of LmACR2 Pentostam in LmACR2 is responsible for the of the reduction of Sb(V) to by LmACR2 be related to the sensitivity of the parasite to The of of LmACR2 in intracellular amastigotes was LmACR2 in was infantum used to to of drug D. PubMed Scopus to Pentostam was in intracellular amastigotes Because is a strain for LmACR2, reductase is is an in sensitivity to Pentostam in intracellular amastigotes LmACR2 in with We that the of LmACR2 be in an This is the first of an enzyme that is in and for of the drug Pentostam in intracellular amastigotes of of LmACR2 in infantum Pentostam infantum the gene D. PubMed Scopus with a with or with LmACR2 with and by a for with the of The of was from The the of with with two of of is by the of drugs PubMed Scopus Pentostam and are the used pentavalent antimony is its as a Pentostam and are of Sb(V) with and Sb(V) and with Sb(III). must be is a in for the treatment of the the parasite in the intracellular amastigote form of is that the Sb(V) in drugs must be reduced to to create the active form of the drug, reduction in the in the or the amastigotes of infantum in have to be resistant to Pentostam intracellular amastigotes in to the that macrophages reduce Pentostam to is by the intracellular amastigotes D. PubMed is by 2004; PubMed Scopus PubMed Scopus and Leishmania species have an that we have shown D. 2004; PubMed Scopus amastigotes of have to reduce and a reduction D. PubMed Scopus that Sb(V) is reduced to in both the and the parasite amastigotes reduce Pentostam D. PubMed Scopus with we an of arsenate and that is an Sb(V) reductase. are to of the enzyme the of purified LmACR2 is that of ScAcr2p, is in the of The of purified LmACR2 be in to the that and Leishmania the of and reductase. are by and reductase PubMed Scopus PubMed Scopus PubMed Scopus PubMed Scopus is PubMed Scopus PubMed Scopus is to that a leishmanial of an arsenate reductase with and for the parasite and is that LmACR2 is a is a LmACR2 in to the ScAcr2p, a arsenate reduction and arsenate with an of PubMed Scopus The and of the enzyme be of Pentostam in amastigotes of Leishmania. Sb(V) is by and a is reduced to is the amastigote by The other of the Sb(V) is the amastigote and reduced to by LmACR2 other such as The of the two to drug the and of in both the and This be in strains of as as in to in drug the of LmACR2 is to be the LmACR2 gene complement the arsenate-sensitive of arsenate reductase of coli and cerevisiae. and of LmACR2 in infantum amastigotes intracellular amastigotes to the amastigotes Sb(V). in we have the of a from with LmACR2 the intracellular amastigotes to the the gene was resistant to that of drug are with of LmACR2 in of the drug to its active of LmACR2 be a of of Sb(V) in LmACR2 be an of drug Leishmania species are distributed worldwide. During its life cycle, the parasite goes through two developmental stages. The promastigote form of the parasite resides in the intestinal tract of the insect vector. The amastigote form of the parasite resides in macrophages and other mononuclear phagocytes in the mammalian host. Between 10 and 15 million people have clinical symptoms of leishmaniasis, and 400,000 new cases are diagnosed each year. Increases in personal and business travel and intervention in regional conflicts such as the last two Iraqi wars have resulted in a significant rise in cases of Leishmania in the United States (1Lockwood D. Nat. Med. 2004; 10: 110Crossref PubMed Scopus (4) The pentavalent drugs Pentostam and are the first line treatment for leishmaniasis, and to drugs is a clinical Med. PubMed Scopus PubMed Scopus To create the active form of the drug, Sb(V) is first reduced to D. PubMed in the are to Pentostam are D. PubMed D. PubMed Scopus and antimony have and are by the 2004; PubMed Scopus PubMed Scopus 2004; PubMed Scopus PubMed Scopus and PubMed Scopus D. PubMed We propose that a of the Sb(V) in Pentostam is reduced by a leishmanial As(V) reductase. are in and and are for to arsenate PubMed Scopus arsenate reductase a Leishmania enzyme related to shown to reduction of arsenate and in 2004; PubMed Scopus is a for a are that a of enzyme and drug sensitivity in Leishmania amastigotes. the arsenate reductase from Saccharomyces arsenate to both in and in PubMed PubMed Scopus PubMed Scopus PubMed Scopus is to the PubMed Scopus and to a PubMed Scopus and as PubMed Scopus Because of its to is to by of We used the sequence to a related Leishmania This LmACR2, was both and Escherichia coli LmACR2 the arsenate-sensitive of the ScACR2 deletion of and a deletion of the coli arsC a arsenate reductase. LmACR2 was purified and The Leishmania enzyme and for arsenate reductase with a of for As(V) and is by and LmACR2 Sb(V) to with a of To a LmACR2 and Pentostam LmACR2 was Leishmania infantum amastigotes from sensitivity to This is the first of an enzyme that is in and for of the drug Pentostam in intracellular Leishmania amastigotes. from and in the form of and from and of Leishmania was for an of The to ScACR2 is in and in The LmACR2, was by by as and as and the and of the for the LmACR2 was by as and as Leishmania LmACR2 was by as and as was first and to the of the in a from The rise to for in for in and for in from coli in a PubMed Scopus with as The arsenate phenotype of was in coli strains or as PubMed Scopus in of arsenate and was from the 15 of and PubMed or in a PubMed or with each of as of of of with The for infantum strain the gene D. PubMed Scopus was used for in in PubMed with 10 as and by of The promastigote used to the line as D. PubMed Scopus with and with infantum a of for and macrophages with of of and was as a D. PubMed Scopus are as of LmACR2 and coli used and purified as PubMed Scopus PubMed Scopus LmACR2 was purified from of coli strain in of with an of was to a of as and the was for an The with in 10 and a of of of and by a through a of was to the The was for and the was a of a with The was with of by with of with the of to LmACR2 was by purified LmACR2 and The from the was to a to with with 10 and with and purified from the by the by the of and PubMed Scopus and LmACR2, of reductase was by a PubMed Scopus The reductase and of was used to that the of the was in the of and as was and as of of LmACR2 by a of for The by was with two of purified of LmACR2 was with the of for the with the used for arsenate as The was with of an of and with was used for of Sb(V) and with as the a of PubMed Scopus Sb(V) Sb(V) a with The of antimony in each was by Sb(V) and of the other The for the of in the of LmACR2, was a of the with of the Sb(V) was the During the of and the used in the reduced from and in the form of and from and of Leishmania was for an of The to ScACR2 is in and in The LmACR2, was by by as and as and the and of the for the LmACR2 was by as and as Leishmania LmACR2 was by as and as was first and to the of the in a from The rise to for in for in and for in Leishmania. from coli in a PubMed Scopus with as The arsenate phenotype of was in coli strains or as PubMed Scopus in of arsenate and was from the 15 of and PubMed or in a PubMed or with each of as of of of with The for infantum strain the gene D. PubMed Scopus was used for in in PubMed with 10 as and by of The promastigote used to the line as D. PubMed Scopus with and with infantum a of for and macrophages with of of and was as a D. PubMed Scopus are as of LmACR2 and coli used and purified as PubMed Scopus PubMed Scopus LmACR2 was purified from of coli strain in of with an of was to a of as and the was for an The with in 10 and a of of of and by a through a of was to the The was for and the was a of a with The was with of by with of with the of to LmACR2 was by purified LmACR2 and The from the was to a to with with 10 and with and purified from the by the by the of and PubMed Scopus and LmACR2, of reductase was by a PubMed Scopus The reductase and of was used to that the of the was in the of and as was and as of of LmACR2 by a of for The by was with two of purified of LmACR2 was with the of for the with the used for arsenate as The was with of an of and with was used for of Sb(V) and with as the a of PubMed Scopus Sb(V) Sb(V) a with The of antimony in each was by Sb(V) and of the other The for the of in the of LmACR2, was a of the with of the Sb(V) was the During the of and the used in the reduced and of Leishmania sequence for an that is in was with and as to a that was and rise to a gene that we LmACR2 LmACR2 and are in and LmACR2 with a of with ScAcr2p, and a of an active the sequence PubMed Scopus and LmACR2 the The two sequence and of LmACR2 the of an of was the in with a from or with the of the arsenate in the coli arsC deletion strain PubMed Scopus LmACR2 of was with gene in the of LmACR2 the As(V) sensitivity of an strain of coli sensitivity to Sb(V) of in LmACR2 is a that was purified by The of purified LmACR2 was by a the sequence of the LmACR2 gene with the and the gene a a of was with a as a PubMed Scopus and of LmACR2 was purified as The of LmACR2 was from its The of the are for and of each of by a of of coli strain with of of and of from an of LmACR2 to reduce arsenate to was by a for and PubMed Scopus PubMed Scopus of arsenate was by LmACR2 in the of the coli for PubMed Scopus and the coli as for LmACR2 the for arsenate was with 10 with The was with both LmACR2 in to the ScAcr2p, and an of PubMed Scopus The of trivalent arsenate reductase was with an of The arsenical and with of and and LmACR2 arsenate reductase in reductase of purified The arsenate reductase of LmACR2 was with and reductase was from the of as purified coli LmACR2 arsenate reductase of purified The arsenate reductase of LmACR2 was with and coli or The of arsenate reductase was from the of each of as The the from two of arsenate reductase by trivalent reductase and of arsenate was in the of of The and of the from the of the of the an the drugs for treatment of such as Pentostam are pentavalent antimonials that must be by reduction to the of LmACR2 to reduce Sb(V) was Pentostam is the of and We used an of trivalent and pentavalent antimonials in the of each species is by PubMed Scopus LmACR2 reduced Sb(V) to in a reductase both and The of Sb(V) reduction a of for Sb(V) and a of the for Sb(V) is for and the is that LmACR2 is a reductase with for Sb(V) Sb(V) to Sb(III). The Sb(V) reductase of LmACR2 was with purified coli and with Sb(V) and by and the of was by as reduction of Sb(V) was for the with of Sb(V) reduction from the of with The line is the to the of LmACR2 Pentostam in LmACR2 is responsible for the of the reduction of Sb(V) to by LmACR2 be related to the sensitivity of the parasite to The of of LmACR2 in intracellular amastigotes was LmACR2 in was infantum used to to of drug D. PubMed Scopus to Pentostam was in intracellular amastigotes Because is a strain for LmACR2, reductase is is an in sensitivity to Pentostam in intracellular amastigotes LmACR2 in with We that the of LmACR2 be in an This is the first of an enzyme that is in and for of the drug Pentostam in intracellular amastigotes of of LmACR2 in infantum Pentostam infantum the gene D. PubMed Scopus with a with or with LmACR2 with and by a for with the of The of was from The the of with with two of and of Leishmania sequence for an that is in was with and as to a that was and rise to a gene that we LmACR2 LmACR2 and are in and LmACR2 with a of with ScAcr2p, and a of an active the sequence PubMed Scopus and LmACR2 the The two sequence and of LmACR2 the of an of was the in with a from or with the of the arsenate in the coli arsC deletion strain PubMed Scopus LmACR2 of was with gene in the of LmACR2 the As(V) sensitivity of an strain of coli sensitivity to Sb(V) of in LmACR2 is a that was purified by The of purified LmACR2 was by a the sequence of the LmACR2 gene with the and the gene a a of was with a as a PubMed Scopus LmACR2 an of LmACR2 to reduce arsenate to was by a for and PubMed Scopus PubMed Scopus of arsenate was by LmACR2 in the of the coli for PubMed Scopus and the coli as for LmACR2 the for arsenate was with 10 with The was with both LmACR2 in to the ScAcr2p, and an of PubMed Scopus The of trivalent arsenate reductase was with an of The arsenical and with of and and LmACR2 arsenate reductase in LmACR2 an the drugs for treatment of such as Pentostam are pentavalent antimonials that must be by reduction to the of LmACR2 to reduce Sb(V) was Pentostam is the of and We used an of trivalent and pentavalent antimonials in the of each species is by PubMed Scopus LmACR2 reduced Sb(V) to in a reductase both and The of Sb(V) reduction a of for Sb(V) and a of the for Sb(V) is for and the is that LmACR2 is a reductase with for Sb(V) of LmACR2 Pentostam in LmACR2 is responsible for the of the reduction of Sb(V) to by LmACR2 be related to the sensitivity of the parasite to The of of LmACR2 in intracellular amastigotes was LmACR2 in was infantum used to to of drug D. PubMed Scopus to Pentostam was in intracellular amastigotes Because is a strain for LmACR2, reductase is is an in sensitivity to Pentostam in intracellular amastigotes LmACR2 in with We that the of LmACR2 be in an This is the first of an enzyme that is in and for of the drug Pentostam in intracellular amastigotes of Leishmania. of is by the of drugs PubMed Scopus Pentostam and are the used pentavalent antimony is its as a Pentostam and are of Sb(V) with and Sb(V) and with Sb(III). must be is a in for the treatment of the the parasite in the intracellular amastigote form of is that the Sb(V) in drugs must be reduced to to create the active form of the drug, reduction in the in the or the amastigotes of infantum in have to be resistant to Pentostam intracellular amastigotes in to the that macrophages reduce Pentostam to is by the intracellular amastigotes D. PubMed is by 2004; PubMed Scopus PubMed Scopus and Leishmania species have an that we have shown D. 2004; PubMed Scopus amastigotes of have to reduce and a reduction D. PubMed Scopus that Sb(V) is reduced to in both the and the parasite amastigotes reduce Pentostam D. PubMed Scopus with we an of arsenate and that is an Sb(V) reductase. are to of the enzyme the of purified LmACR2 is that of ScAcr2p, is in the of The of purified LmACR2 be in to the that and Leishmania the of and reductase. are by and reductase PubMed Scopus PubMed Scopus PubMed Scopus PubMed Scopus is PubMed Scopus PubMed Scopus is to that a leishmanial of an arsenate reductase with and for the parasite and is that LmACR2 is a is a LmACR2 in to the ScAcr2p, a arsenate reduction and arsenate with an of PubMed Scopus The and of the enzyme be the of LmACR2 is to be the LmACR2 gene complement the arsenate-sensitive of arsenate reductase of coli and cerevisiae. and of LmACR2 in infantum amastigotes intracellular amastigotes to the amastigotes Sb(V). in we have the of a from with LmACR2 the intracellular amastigotes to the the gene was resistant to that of drug are with of LmACR2 in of the drug to its active of LmACR2 be a of of Sb(V) in LmACR2 be an of drug of is by the of drugs PubMed Scopus Pentostam and are the used pentavalent antimony is its as a Pentostam and are of Sb(V) with and Sb(V) and with Sb(III). must be is a in for the treatment of the the parasite in the intracellular amastigote form of is that the Sb(V) in drugs must be reduced to to create the active form of the drug, reduction in the in the or the amastigotes of infantum in have to be resistant to Pentostam intracellular amastigotes in to the that macrophages reduce Pentostam to is by the intracellular amastigotes D. PubMed is by 2004; PubMed Scopus PubMed Scopus and Leishmania species have an that we have shown D. 2004; PubMed Scopus amastigotes of have to reduce and a reduction D. PubMed Scopus that Sb(V) is reduced to in both the and the parasite amastigotes reduce Pentostam D. PubMed Scopus with we an of arsenate and that is an Sb(V) reductase. are to of the enzyme the of purified LmACR2 is that of ScAcr2p, is in the of The of purified LmACR2 be in to the that and Leishmania the of and reductase. are by and reductase PubMed Scopus PubMed Scopus PubMed Scopus PubMed Scopus is PubMed Scopus PubMed Scopus is to that a leishmanial of an arsenate reductase with and for the parasite and is that LmACR2 is a is a LmACR2 in to the ScAcr2p, a arsenate reduction and arsenate with an of PubMed Scopus The and of the enzyme be the of LmACR2 is to be the LmACR2 gene complement the arsenate-sensitive of arsenate reductase of coli and cerevisiae. and of LmACR2 in infantum amastigotes intracellular amastigotes to the amastigotes Sb(V). in we have the of a from with LmACR2 the intracellular amastigotes to the the gene was resistant to that of drug are with of LmACR2 in of the drug to its active of LmACR2 be a of of Sb(V) in LmACR2 be an of drug We for with Leishmania and for and with

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.