[大鼠唾液腺、脑、肝、肾唾液酸酶的性质]。

A Sato
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引用次数: 0

摘要

唾液酸酶(SD, neuroaminidase, EC 3.18)是一种从糖蛋白、糖脂和低聚糖中释放末端唾液酸残基的酶。在本研究中,我们报道了该酶在大鼠唾液腺、脑、肝和肾中的一些特征。用合成底物4-methylumbelliferyl- α - d - n -acetylneuraminic acid (4MU-AcNeu)荧光法测定SD活性。1. 组织匀浆中SD活性在pH值为4.0时在下颌骨腺(SMG)最大,在pH值为4.5时在舌下腺(SLG)和腮腺(PG)最大,其值与脑、肝和肾的活性值非常相似。SMG酶活性最高,PG次之,SLG次之。然而,这些腺体的SD活性低于脑、肝和肾。2. 细胞分离显示SD在唾液腺的细胞内分布以溶酶体部分为主,与在脑、肝、肾的分布相似。3.SMG溶酶体SD的最适pH值为4.0,SLG和PG溶酶体SD的最适pH值为4.5,所有唾液腺溶酶体SD的4MU-AcNeu的Km值约为0.09 mM,而SMG和PG溶酶体SD的最适pH值为5.5,4MU-AcNeu的Km值为0.25mM。4. 热稳定性研究表明,SD在唾液腺和其他器官中非常不稳定,可溶性SD比溶酶体SD更不稳定。5. PG、脑和肝脏溶酶体SD倾向于被低浓度Ca2+激活。Ca2+对SMG、PG和肝脏的可溶性SD也有这种影响。此外,SMG中的可溶性SD被低浓度的Mg2+轻微活化。Cu2+和Hg2+对溶酶体和可溶性SD均有明显的抑制作用。6. 通过Sephacryl S-400凝胶过滤,估计SMG和SLG溶酶体的分子量(MW)为52万,PG为46万道尔顿,脑、肝、肾溶酶体的分子量为35万~ 46万道尔顿。另一方面,通过Sephadex G-200凝胶过滤,SMG和PG中可溶性SD的分子量分别为68,000和46,000。脑和肝脏中可溶性SD的分子量与PG中相应酶的分子量非常相似。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Properties of sialidases from salivary glands, brain, liver and kidney of the rat].

Sialidase (SD, neuraminidase, EC 3.2.1 18) is an enzyme which releases terminal sialic acid residues from glycoproteins, glycolipids nad oligosaccharides. In this study, we report some characteristics of this enzyme in rat salivary glands, brain, liver and kidney. SD activity was measured fluorometrically by using the synthetic substrate 4-methylumbelliferyl-alpha-D-N-acetylneuraminic acid (4MU-AcNeu). 1. SD activity in tissue homogenates was maximal at pH 4.0 in the submandibular gland (SMG), and at pH 4.5 in both the sublingual gland (SLG) and parotid gland (PG), the values being very similar to those for the activity in brain, liver and kidney. Enzyme activity was highest in SMG, followed by PG and SLG. However, SD activity was lower in these glands than in brain, liver and kidney. 2. Cell fractionation showed that the intracellular distribution of SD in the salivary glands was mainly in the lysosomal fraction, similar to its distribution in brain, liver or kidney. 3. The pH optimum of lysosomal SD was 4.0 in the SMG, and 4.5 in both SLG and PG. The Km value for 4MU-AcNeu of lysosomal SD from all salivary glands was about 0.09 mM. On the other hand, soluble SD in SMG and PG extracts had its pH optimum at 5.5, and a Km value of 0.25mM for 4MU-AcNeu. 4. Studies on heat stability showed SD in the salivary glands and other organs to be very labile, with soluble SD being more labile than lysosomal SD. 5. Lysosomal SD in the PG, brain and liver was inclined to be activated by Ca2+ at low concentrations. Such an effect of Ca2+ was also seen with soluble SD in the SMG, PG and liver. In addition, soluble SD in the SMG was slightly activated by low concentrations of Mg2+. Both Cu2+ and Hg2+ caused a marked inhibition of lysosomal and soluble SD in all organs. 6. By means of Sephacryl S-400 gel filtration, the molecular weight (MW) of lysosomal SD was estimated to be 520,000 in both SMG and SLG, and 460,000 daltons in the PG. The MW of the enzyme in brain, liver and kidney ranged from 350,000 to 460,000. On the other hand, the MW's of soluble SD in SMG and PG were estimated to be 68,000 and 46,000, respectively, by Sephadex G-200 gel filtration. The MW's of soluble SD in brain and liver were very similar to the MW of the corresponding enzyme in the PG.(ABSTRACT TRUNCATED AT 400 WORDS)

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