Purification and characterization of arylalkylamine N-acetyltransferase from cockroach testicular organs

Ichihara, Naoyuki; Okada, Masato; Nakagawa, Hachiro; Takeda, Makio

doi:10.1016/s0965-1748(96)00091-4

Cited by 20 publications

(22 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Based on the physiological requirements of insects, all of these proposed physiological functions regarding insect aaNATs are reasonable predictions. aaNAT proteins have been studied in Drosophila melanogaster (Amherd et al, 2000; Hintermann et al, 1996) and Periplaneta americana (Ichihara et al, 1997; Ichihara et al, 2001). And, recent studies dealing with an aaNAT from Bombyx mori clearly established its function in adult cuticle sclerotization through the production of N -acetyldopamine, a key cuticle protein crosslinking precursor (Dai et al, 2010; Zhan et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Identification and characterization of two arylalkylamine N-acetyltransferases in the yellow fever mosquito, Aedes aegypti

Mehere

Han

Christensen

et al. 2011

Insect Biochemistry and Molecular Biology

View full text Add to dashboard Cite

In this study we provide a molecular and biochemical identification of two arylalkylamine N-acetyltransferases (aaNAT) from Aedes aegypti mosquitoes. N-acetyldopamine, the enzyme product of aaNAT, was detected in Ae. aegypti, indicating the presence of an aaNAT in this mosquito. A BLAST search of the Ae. aegypti genome, using sequence information from an activity-verified Drosophila aaNAT, identified thirteen putative aaNAT sequences sharing 13-48% sequence identity with the Drosophila enzyme. Eight of the thirteen putative aaNAT proteins were expressed using a bacterial expression system. Screening of purified recombinant proteins against 5-hydroxytryptamine, dopamine, methoxytryptamine, norepinephrine, octopamine, tryptamine, and tyramine substrates, established that two of the putative aaNATs are active to the tested arylalkylamines. We therefore named them aaNAT1 and 2, respectively. Analysis of the transcriptional profiles of the two aaNAT genes from Ae. aegypti revealed that aaNAT1 is more abundant in the whole body of larvae and pupae, and aaNAT2 is more abundant in the head of adult mosquitoes. Based on their substrate and transcriptional profiles, together with previous reports from other insects, we suggest that the two aaNATs play diverse roles in Ae. aegypti, with aaNAT1 primarily involved in sclerotization and aaNAT2 mainly in neurotransmitter inactivation. Our data provide a beginning to a more comprehensive understanding of the biochemistry and physiology of aaNATs from the Ae. aegypti and serve as a reference for studying the aaNAT family of proteins from other insect species.

show abstract

Section: Introductionmentioning

confidence: 99%

Identification and characterization of two arylalkylamine N-acetyltransferases in the yellow fever mosquito, Aedes aegypti

Mehere

Han

Christensen

et al. 2011

Insect Biochemistry and Molecular Biology

View full text Add to dashboard Cite

show abstract

“…The asterisks indicate cysteine residues which probably form structurally important disulfide bonds. Boldface type represents amino acid sequences derived from peptide I and II of NAT purified by Ichihara et al (1997). Motifs A and B are underlined (see text).…”

Section: Resultsmentioning

confidence: 99%

“…Degenerate oligonucleotide primers were designed corresponding to the amino acid sequences 1-7 and 4-10 from peptide I (FFFLEEPLNI) and the 4-9 amino acid sequence from peptide II (KLLGENGDE) previously obtained by chromatographic separation (Ichihara et al, 1997). These primers are for sense (5'-TTY TTY TTY YTI GAR GAR CC-3' or 5'-Y YTI GAR GAR CCI YTI AAY A-3') and antisense (5'-TC RTC KCC RTT YTC NCC-3', where N is any base; Y, C/T; R, A/G and I, inosine) and the ds cDNA was used as template.…”

Section: Partial Characterization Of N-acetyltransferase From the Tesmentioning

confidence: 99%

“…After cloning into pT7Blue T-vector (Novagen, Madison, WI), the sequence of ten independent clones was determined. The sequences contained an ORF that extended beyond the boundary of sequence homologies with peptides previously described by Ichihara et al (1997). The nucleotide sequences obtained from all the clones were simi-…”

Section: Partial Characterization Of N-acetyltransferase From the Tesmentioning

confidence: 99%

“…The predicted size of NAT was 28 kDa with a pI of 6.0. Ichihara et al (1997) determined partial amino acid sequences (referred to as peptide I and II in this report) derived from proteolytic fragments of the NAT thus obtained. Using these sequences for degenerate primers, we cloned a full-length cDNA encoding an N-acetyltransferase.…”

Section: August 2005mentioning

confidence: 99%

See 2 more Smart Citations

Molecular cloning of a cDNA encoding arylalkylamineN-acetyltransferase from the testicular system ofPeriplaneta americana: Primary protein structure and expression analysis

Bembenek

Sakamoto

Takeda

2005

Arch. Insect Biochem. Physiol.

Self Cite

View full text Add to dashboard Cite

DNA encoding a fragment of putative arylalkylamine N-acetyltransferase (NAT) of the American cockroach, Periplaneta americana, was amplified by PCR with degenerate primers based on the two peptides previously purified from the testicular system of this species. A full clone was obtained by RACE-PCR. The clone consisted of 89 bp 5'-UTR, 753-bp open reading frame and 712-bp 3'-UTR. The amino acid sequence of 251 residues deduced from this ORF corresponded to the predicted molecular mass of 28.5 kDa. The predicted amino acid sequence had an overall identity of 35% with NAT1 and 27% with NAT2 of Drosophila melanogaster, respectively. Structural analysis revealed that NAT from P. americana contained two motifs characteristic of the NAT superfamily and three conserved regions (C/c-1, D/c-1, D/c-2) distinguishing aaNAT subfamily. Northern blot analysis showed that the mRNA of approximately 1.5 kb was transcribed at a high level in the testicular system, and corresponded to the length of the cDNA, i.e., 1,554 bp. Significant levels of NAT transcript were also detected in the midgut, ovary and the accessory glands and at much lower levels in the fat body and brain. Southern blot analysis suggested the presence of a single copy of the cloned gene in the genome.

show abstract

Arylalkalamine N‐acetyltransferase‐1 acts on a secondary amine in the yellow fever mosquito, Aedes aegypti

et al. 2022

View full text Add to dashboard Cite

Arylalkylamine N-acetyltransferase (aaNAT) in Aedes aegypti is primarily involved in cuticle pigmentation and formation. The reported arylalkylamine substrates are all primary amines. In this study, we report a novel substrate, a secondary amine, of Ae. aegypti aaNAT1. The recombinant aaNAT1 protein exhibited high activity to a secondary amine, epinephrine, which has not been reported for any aaNATs previously. Structure-activity relationship study demonstrated that aaNAT1 has an epinephrine-binding site, and molecular docking and dynamic simulation showed that epinephrine is quite stable in the active cavity. Further functional studies demonstrated that epinephrine affected mosquito fecundity, egg hatching and development. The new biochemical function of aaNAT1 in metabolizing epinephrine could reduce some negative effects of the compound in the mosquito.

show abstract

Purification and characterization of arylalkylamine N-acetyltransferase from cockroach testicular organs

Cited by 20 publications

References 17 publications

Identification and characterization of two arylalkylamine N-acetyltransferases in the yellow fever mosquito, Aedes aegypti

Identification and characterization of two arylalkylamine N-acetyltransferases in the yellow fever mosquito, Aedes aegypti

Molecular cloning of a cDNA encoding arylalkylamineN-acetyltransferase from the testicular system ofPeriplaneta americana: Primary protein structure and expression analysis

Arylalkalamine N‐acetyltransferase‐1 acts on a secondary amine in the yellow fever mosquito, Aedes aegypti

Contact Info

Product

Resources

About