Department of Biochemistry, Virginia Tech, Blacksburg, Virginia, United States of America.
PLoS One. 2011 Jan 24;6(1):e16124. doi: 10.1371/journal.pone.0016124.
One protein in Aedes aegypti, classified into the aromatic amino acid decarboxylase (AAAD) family based on extremely high sequence homology (∼70%) with dopa decarboxylase (Ddc), was biochemically investigated. Our data revealed that this predicted AAAD protein use L-dopa as a substrate, as does Ddc, but it catalyzes the production of 3,4-dihydroxylphenylacetaldehyde (DHPAA) directly from L-dopa and apparently has nothing to do with the production of any aromatic amine. The protein is therefore named DHPAA synthase. This subsequently led to the identification of the same enzyme in Drosophila melanogaster, Anopheles gambiae and Culex quinquefasciatus by an initial prediction of putative DHPAA synthase based on sequence homology and subsequent verification of DHPAA synthase identity through protein expression and activity assays. DHPAA is highly toxic because its aldehyde group readily reacts with the primary amino groups of proteins, leading to protein crosslinking and inactivation. It has previously been demonstrated by several research groups that Drosophila DHPAA synthase was expressed in tissues that produce cuticle materials and apparent defects in regions of colorless, flexible cuticular structures have been observed in its gene mutants. The presence of free amino groups in proteins, the high reactivity of DHPAA with the free amino groups, and the genetically ascertained function of the Drosophila DHPAA synthase in the formation of colorless, flexible cuticle, when taken together, suggest that mosquito and Drosophila DHPAA synthases are involved in the formation of flexible cuticle through their reactive DHPAA-mediated protein crosslinking reactions. Our data illustrate how a seemingly highly toxic pathway can serve for an important physiological function in insects.
在埃及伊蚊中发现了一种蛋白,根据与多巴脱羧酶(Ddc)极高的序列同源性(约 70%),将其归类为芳香族氨基酸脱羧酶(AAAD)家族。我们的研究数据表明,这种预测的 AAAD 蛋白像 Ddc 一样使用 L-多巴作为底物,但它直接从 L-多巴催化生成 3,4-二羟基苯乙醛(DHPAA),显然与任何芳香胺的生成无关。因此,该蛋白被命名为 DHPAA 合酶。这随后通过基于序列同源性的初始预测和随后通过蛋白表达和活性测定来验证 DHPAA 合酶的身份,在果蝇、冈比亚按蚊和致倦库蚊中鉴定到了相同的酶。DHPAA 具有高度毒性,因为其醛基很容易与蛋白质的一级氨基反应,导致蛋白质交联和失活。几个研究小组已经证明,果蝇 DHPAA 合酶在产生角质材料的组织中表达,并且在其基因突变体中观察到无色、柔韧的角质结构区域的明显缺陷。蛋白质中存在游离氨基基团、DHPAA 与游离氨基基团的高反应性,以及果蝇 DHPAA 合酶在无色、柔韧角质形成中的遗传确定功能,这一切都表明,蚊子和果蝇的 DHPAA 合酶通过其反应性的 DHPAA 介导的蛋白交联反应参与柔韧角质的形成。我们的数据说明了看似高度毒性的途径如何在昆虫中发挥重要的生理功能。