Acetylcholinesterase from the Skeletal Muscle of the Lamprey Petromyzon marinus Exists in Globular and Asymmetric Forms
Acetylcholinesterase from the Skeletal Muscle of the Lamprey Petromyzon marinus Exists in Globular and Asymmetric Forms
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来自七鳃鳗骨骼肌的乙酰胆碱酯酶以球状和不对称形式存在
DOI：
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发表时间：
1987
影响因子：
4.7
通讯作者：
Maureen L. Pezzementi
中科院分区：
文献类型：
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作者：
Leo Pezzementi;Ellen J. Reinheimer;Maureen L. Pezzementi
摘要：为了获得有关乙酰胆碱酯酶(AChE)进化的信息，我们对原始脊椎动物七鳃鳗骨骼肌中的乙酰胆碱酯酶进行了研究。我们发现七鳃鳗肌肉的胆碱酯酶活性是由AChE引起的，而不是假性胆碱酯酶，该酶被1，5-二(4-烯丙基二甲基苯基)戊烷-3-酮(BW284C51)抑制，但不被四单异丙基焦磷酸四胺(iso-OmpA)或乙丙嗪抑制。此外，该酶对乙酰硫代胆碱有较高的亲和力，并受高浓度底物的抑制。用刀豆蛋白A-琼脂糖沉淀AChE，发现很大一部分AChE是糖蛋白。AChE的最佳提取条件是在含高盐洗涤剂的缓冲液中提取；在没有盐和/或洗涤剂的缓冲液中提取少量的酶。这些数据表明，存在球状和不对称形式的AChE。在蔗糖梯度上，在含高盐洗涤剂的缓冲液中提取的酶沉淀为对应于G4的宽峰；此外，通常有对应于A12的峰。连续提取AChE结合速度沉降法解决了较小形式的AChE，揭示了G1、G2、G4、A4、A8和A12形式的AChE可从肌肉中获得。通过高盐沉淀和胶原酶消化证实了这两种形式的一致性。在低离子强度的缓冲液中，不对称形式的AChE被沉淀下来，胶原酶消化使其沉降系数向较高的值移动。我们从后口脊椎动物和无脊椎动物中AChE的进化以及不对称形式的AChE对肌肉功能的生理学意义的角度讨论了这些结果。
Abstract: To obtain information about the evolution of acetylcholinesterase (AChE), we undertook a study of the enzyme from the skeletal muscle of the lamprey Petromyzon marinus, a primitive vertebrate. We found that the cholines‐terase activity of lamprey muscle is due to AChE, not pseudocholinesterase; the enzyme was inhibited by 1,5‐bis(4‐allyldimethylammonium phenyl) pentane‐3‐one (BW284C51), but not by tetramonoisopropyl pyrophos‐phortetramide (iso‐OMPA) or ethopropazine. Also, the enzyme had a high affinity for acetylthiocholine and was inhibited by high concentrations of substrate. A large fraction of the AChE was found to be glycoprotein, since it was precipitated by concanavalin A‐agarose. Optimal extraction of AChE was obtained in a high‐salt detergent‐containing buffer; fractional amounts of enzyme were extracted in buffers lacking salt and/or detergent. These data suggest that globular and asymmetric forms of AChE are present. On sucrose gradients, enzyme that was extracted in high‐salt detergent‐containing buffer sedimented as a broad peak of activity corresponding to G4; additionally, there was usually a peak corresponding to A12. Sequential extraction of AChE in conjunction with velocity sedimentation resolved minor forms of AChE and revealed that the G1, G2, G4, A4, A8, and A12 forms of AChE cpuld be obtained from the muscle. The identity of the forms was confirmed through high‐salt precipitation and collagenase digestion. The asymmetric forms of AChE were precipitated in low ionic strength buffer, and their sedimentation coefficients were shifted to higher values by collagenase digestion. We discuss these results in terms of the evolution of AChE in deuterostome vertebrates and invertebrates and in terms of the physiological significance of asymmetric forms of AChE for muscle function.
人红细胞乙酰胆碱酯酶的结构。
DOI：
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发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Rosenberry,TL;Scoggin,DM
通讯作者：
Scoggin,DM
来自 Torpedo 的不对称 (17 13) S 乙酰胆碱酯酶的结构表征。
DOI：
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发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lee,SL;Taylor,P
通讯作者：
Taylor,P
来自 Torpedo 的乙酰胆碱酯酶的不对称 (17 13) S 形式的结构表征。
DOI：
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发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lee,SL;Heinemann,S;Taylor,P
通讯作者：
Taylor,P