CORE--Solution properties of alternatively spliced proteins / Core 1

这是项目“从基因组序列到蛋白质结构和功能”的一个新的核心组成部分。目标是确定选定用于结构研究的蛋白质的溶液性质。预计在下一阶段研究的许多人类蛋白质将需要鼓励才能折叠成其功能构象，并且许多选择性剪接的蛋白质可能没有折叠的功能结构。虽然它是一个很小的核心，但它对成功是必不可少的，因为它将确保适当地重视确定我们蛋白质的溶液性质。PL在研究蛋白质折叠和处理蛋白质溶液性质方面的丰富经验将在整个项目中发挥重要作用。将使用一套标准的生物物理技术： (1)圆二色谱，以获得每个蛋白质折叠状态的初步指示。 (2)一维和二维核磁共振谱，以提供每种蛋白质的有序程度和水平的准确信息。 (3)。MALDI-TOF质谱仪，提供精确的相对分子质量和纯度信息。 (4)。动态光散射，用于初步识别可能干扰的大集合体 结晶，并建立蛋白质单分散的条件。 (5)除动态光散射外，还可用于尺寸排除层析，以评估样品的均质性、聚集性和低聚状态。 (6)。超速离心法，准确测定每种合适蛋白质的寡聚状态。预计在某些情况下，不同的选择性剪接蛋白版本会有不同的低聚状态。

This is a new core component of the program project 'From Genomic Sequences to Protein Structure and function'. The goal is to determine the solution properties of proteins selected for structural studies. It is expected that many of the human proteins studied in the next phase will require encouragement to fold into their functional conformation, and that a number of the alternatively spliced proteins may not have a folded functional structure. Although it is a small core, it is essential for success, as it will ensure that proper emphasis is placed on determining the solution properties of our proteins. The Pl's extensive experience in studying protein folding, and dealing with solution properties of proteins will be of value throughout the project. A set of standard biophysical techniques wilt be used: (1) Circular dichroism, to obtain an initial indication of the folding state of each protein. (2) One and two-dimensional NMR spectra, to provide precise information of the extent and level of ordering of each protein. (3). MALDI-TOF mass spectroscopy, to provide precise molecular weight and purity information. (4). Dynamic Light Scattering, for initial identification of large aggregates that might interfere with crystallization, and for establishing conditions in which the protein is monodisperse. (5) Size exclusion chromatography, used in addition to Dynamic Light Scattering, to assess the homogeneity, aggregation, and oligomeric state of the sample. (6). Ultracentrifugation, to determine accurately the oligomeric state of each suitable protein. It is expected that in some cases, different alternative splice protein versions will have different oligomeric states.