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斑纹强棱蜥(爬行纲:角蜥科)胃肠道几丁质酶的特性分析

Characterization of gastrointestinal chitinase in the lizard Sceloporus undulatus garmani (Reptilia: Phrynosomatidae).

作者信息

Marsh R S, Moe C, Lomneth R B, Fawcett J D, Place A

机构信息

Department of Biology, University of Nebraska at Omaha, 6001 Dodge Street, Omaha, NE 68182, USA.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2001 Apr;128(4):675-82. doi: 10.1016/s1096-4959(00)00364-x.

Abstract

Most studies on chitinase activity in lizards have been concerned with Palaearctic (European) and Laurasian (Middle Eastern and Asian) taxa. Several genera of Old World lizards, Anguis, Uromastix, Chamaeleo and Lacerta, have been shown to possess chitinolytic activity. To date, only one New World lizard, Anolis carolinensis, has been reported to exhibit chitinolytic activity. In the present study, chitinase activity was characterized in a second New World taxon, Sceloporus undulatus garmani, a New World, phrynosomatid lizard. Chitinolytic activity was measured by incubating tissue extracts with a radioactive chitin substrate, acetyl-[H3]chitin and determining acid soluble radioactivity as an estimate for chitin hydrolysis. Chitinolytic activity was present in stomach, small intestine and pancreas extracts, with the stomach and pancreas having the highest specific activities. Chitinolytic activity was higher at pH 4.5 than at pH 7.5. The stomach chitinase is immunologically similar to the gastric chitinase previously described for rainbow trout. Western blot analysis showed anti-chitinase cross-reactivity in the extracts of the stomach, but no cross-reactivity in the pancreatic or intestinal extracts, suggesting different isoforms of chitinase. There was no detected lysozyme activity (less than 0.01 mg/ml lysozyme) present in the extracts of the stomach, small intestine and pancreas. The localization of chitinolytic activity in S. u. garmani is in agreement with earlier reptilian reports on the distribution of chitinase.

摘要

大多数关于蜥蜴几丁质酶活性的研究都集中在古北区(欧洲)和劳亚古陆区(中东和亚洲)的类群上。已证实旧大陆蜥蜴的几个属,如盲蛇属、沙蜥属、避役属和蜥蜴属,具有几丁质分解活性。迄今为止,仅报道过一种新大陆蜥蜴,即卡罗来纳安乐蜥,具有几丁质分解活性。在本研究中,对新大陆的另一个类群——饰纹强棱蜥(Sceloporus undulatus garmani),一种新大陆的角蜥科蜥蜴,进行了几丁质酶活性分析。通过将组织提取物与放射性几丁质底物乙酰 - [H3]几丁质一起孵育,并测定酸溶性放射性来衡量几丁质分解活性,以此作为几丁质水解的估计值。几丁质分解活性存在于胃、小肠和胰腺提取物中,胃和胰腺的比活性最高。几丁质分解活性在pH 4.5时比在pH 7.5时更高。胃几丁质酶在免疫学上与先前描述的虹鳟鱼胃几丁质酶相似。蛋白质印迹分析显示,胃提取物中有抗几丁质酶交叉反应,但胰腺或肠道提取物中没有交叉反应,这表明几丁质酶存在不同的同工型。在胃、小肠和胰腺提取物中未检测到溶菌酶活性(低于0.01 mg/ml溶菌酶)。饰纹强棱蜥中几丁质分解活性的定位与早期关于几丁质酶分布的爬行动物报告一致。

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