Chen S, Wetzel R
Graduate School of Medicine, University of Tennessee Medical Center, Knoxville, Tennessee 37920, USA.
Protein Sci. 2001 Apr;10(4):887-91. doi: 10.1110/ps.42301.
A method is described for dissolving and disaggregating chemically synthesized polyglutamine peptides. Polyglutamine peptides longer than about Q20 have been reported to be insoluble in water, but dissolution in--and evaporation from--a mixture of trifluoroacetic acid and hexafluoroisopropanol converts polyglutamine peptides up to at least Q44 to a form readily soluble in aqueous buffers. This procedure also has a dramatic effect on peptides which appear to be completely soluble in water, by removing traces of aggregate that seed aggregation. The protocol makes possible solution studies-including in vitro aggregation experiments--on polyglutamine peptides with repeat lengths associated with increased risk of Huntington's Disease and other expanded CAG repeat diseases. It may also be useful in conducting reproducible, quantitative aggregation studies on other polypeptides.
描述了一种溶解和分解化学合成的聚谷氨酰胺肽的方法。据报道，长度超过约Q20的聚谷氨酰胺肽不溶于水，但在三氟乙酸和六氟异丙醇的混合物中溶解并蒸发后，可将至少长达Q44的聚谷氨酰胺肽转化为易溶于水性缓冲液的形式。该方法还通过去除引发聚集的微量聚集体，对似乎完全可溶于水的肽产生显著影响。该方案使得对与亨廷顿舞蹈症及其他CAG重复序列扩展疾病风险增加相关的重复长度的聚谷氨酰胺肽进行溶液研究（包括体外聚集实验）成为可能。它也可能有助于对其他多肽进行可重复的定量聚集研究。
