Revealing the Amylase Interactome in Whole Saliva Using Proteomic Approaches
Revealing the Amylase Interactome in Whole Saliva Using Proteomic Approaches
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DOI：
10.1155/2018/6346954
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Siqueira, Walter Luiz
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文献类型：
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作者：
Crosara, Karla Tonelli Bicalho;Zuanazzi, David;Siqueira, Walter Luiz
了解唾液中存在的蛋白质及其分离时的功能不足以描述它们在口腔中的真实的作用。由于蛋白质-蛋白质相互作用，大分子可能发生结构变化，导致功能调节或修饰。除了淀粉酶在碳水化合物分解中的功能之外，淀粉酶还可以延迟蛋白质伴侣（例如，组胺素1）复合时。由于其生物化学特性和唾液中的高丰度，淀粉酶可能与作为生物载体的几种蛋白质相互作用。本研究的重点是确定淀粉酶和其他蛋白质之间的相互作用，发现在整个唾液（WS）使用蛋白质组学的方法。使用亲和色谱法，然后进行凝胶电泳法、十二烷基硫酸钠和天然、胰蛋白酶溶液内和凝胶内消化以及质谱法。我们鉴定了66个与WS中淀粉酶相互作用的蛋白质。鉴定的蛋白质的特征表明，酸性（pI < 6.8）和低分子量（MW < 56 kDa）蛋白质在淀粉酶复合物形成过程中具有偏好性。大多数已鉴定的蛋白质具有与宿主保护相关的生物学功能。利用STRING数据库构建了一个新的蛋白质-淀粉酶网络。进一步的研究是必要的，以调查所确定的淀粉酶相互作用的个性。这些观察结果为淀粉酶尚未完全表征的生物学功能的更全面研究开辟了途径。
Understanding proteins present in saliva and their function when isolated is not enough to describe their real role in the mouth. Due to protein-protein interactions, structural changes may occur in macromolecules leading to functional modulation or modification. Besides amylase's function in carbohydrate breakdown, amylase can delay proteolytic degradation of protein partners (e.g., histatin 1) when complexed. Due to its biochemical characteristics and high abundance in saliva, amylase probably interacts with several proteins acting as a biological carrier. This study focused on identifying interactions between amylase and other proteins found in whole saliva (WS) using proteomic approaches. Affinity chromatography was used, followed by gel electrophoresis methods, sodium dodecyl sulfate and native, tryptic in-solution and in-gel digestion, and mass spectrometry. We identified 66 proteins that interact with amylase in WS. Characterization of the identified proteins suggests that acidic (pI < 6.8) and low molecular weight (MW < 56 kDa) proteins have preference during amylase complex formation. Most of the identified proteins present biological functions related to host protection. A new protein-amylase network was constructed using the STRING database. Further studies are necessary to investigate individualities of the identified amylase interactors. These observations open avenues for more comprehensive studies on not yet fully characterized biological function of amylase.