RUI: Conformation Analysis of Proteinaceous Bioadhesives

9513390 Olivieri本研究旨在阐明与蛋白质结合固体表面的物理化学机制相关的分子结构。 为了实现本发明的具体目的，有必要获得有助于计算已知生物粘合剂的三维结构的数据。 具体地，从蓝海贻贝（Mytilus edulis）中提取的MAP将提供模型化合物。 MAP由75（85）个重复的十肽单元组成，其一级序列为NH 2-A-K-P-S-Y-HYP-HYP-T-DOPA-K-COOH（Waite等人，1985年）。 由于MAP在界面处是活性的，因此研究其吸附到表面时的行为是重要的。 固态技术，如多重衰减内反射红外（MAIR-IR）光谱，椭圆偏振，接触角分析，和溶液状态核磁共振（NMR）光谱，已经给出了一些关于这些肽的能力的结构信息，但还没有被用来充分发挥其潜力。 除了结晶努力之外，这些技术还将用于研究十聚体重复单元以及其他MAP蛋白片段（例如，+ NH 3-脯氨酸-丝氨酸-酪氨酸-羟脯氨酸-羟脯氨酸-苏氨酸-酪氨酸-赖氨酸-丙氨酸-赖氨酸-脯氨酸-丝氨酸-酪氨酸-羟脯氨酸-COOH，和+NH 3-脯氨酸-丝氨酸-酪氨酸-羟脯氨酸-羟脯氨酸-苏氨酸-3，4-二羟基苯丙氨酸-赖氨酸-丙氨酸-赖氨酸-脯氨酸-丝氨酸-酪氨酸-羟脯氨酸-COOH）。 将收集固态和溶液态MAP肽的数据，并将信息汇编以创建肽的分子模型。 结构解析的第一步将是使用NMR数据来创建溶液状态模型。 各种固态技术将被用来表征负责生物粘附过程的分子变化。 PI致力于将联合收割机研究与教学相结合。 她把她的教学责任非常认真，并致力于为学生提供一个学习环境，是彻底的，但邀请。 PI在D 'YC教授的各种课程反映了她的多学科背景（计算机导论，统计学，生物化学，普通化学I-II，学生研究I-II和健康科学化学II）。 她为生物化学课程准备了80多页的讲义，为健康科学实验室的化学共同撰写了一本手册，并正在为生物化学实验室编写一本手册。 为了增加学生的可用性，她举办学习班，并分发她的家庭电话号码。 一些学生将部分成功归功于分发的讲义、批改的家庭作业和口头报告。 将开发一门生物材料课程，介绍植入学，并描述用于特定生物应用的材料的选择和表征方法。 PI已经为她的生物化学实验室课程开发了几个新的实验室练习，并将准备结合联合收割机科学学科的额外练习。 ***

9513390 Olivieri This study seeks to elucidate the molecular structure(s) associated with the physical-chemical mechanism(s) whereby proteins bind to solid surfaces. To achieve the specific aim of the current proposal it is necessary to obtain data that will assist in the calculation of the three dimensional structures of known bioadhesives. Specifically, MAP extracted from the Blue Sea Mussel, (Mytilus edulis) will provide the model compounds. MAP is composed of 75(85 repeating decapeptide units with the primary sequence NH2-A-K-P-S-Y-HYP-HYP-T-DOPA-K-COOH (Waite et al., 1985). Since MAP is active at interfaces, it is therefore important to study its behavior while adsorbed to a surface. Solid-state techniques, such as multiple attenuated internal reflectance infrared (MAIR-IR) spectroscopy, ellipsometry, contact angle analysis, and solution-state nuclear magnetic resonance (NMR) spectroscopy, already have given some structural information about these peptides' abilities, but have not been used to their fullest potential. These techniques, in addition to crystallization efforts, will be employed to study the decameric repeat unit in addition to other MAP protein fragments (e.g., +NH3-proline-serine-tyrosine-hydroxyproline-hydroxyproline-thereonine-tyrosine-lysine-alanine-lysine-proline-serine-tryosine-hydroxyproline-COOH, and +NH3-proline-serine-tyrosine-hydroxyproline-hydroxyproline-threonine-3,4-dihydroxyphenylalanine-lysine-alanine-lysine-proline-serine-tyrosine-hydroxyproline-COOH). Data will be collected for the MAP peptides in both the solid-and solution-states, with the information compiled to create molecular models of the peptides. The first step of structure elucidation will be to use NMR data to create solution-state models. The variety of solid-state techniques would then be used to characterize the molecular changes that are responsible for the bioadhesion process. %%% The PI has worked to combine research with teaching. She takes her teaching responsibilities very s eriously, and is dedicated toward providing a learning environment for students that is thorough, yet inviting. The variety of courses the PI has taught at D'YC reflects her multi-disciplinary background (Introduction to Computers, Statistics, Biochemistry, General Chemistry I-II, Student Research I-II, and Chemistry for the Health Sciences II). She has prepared a 80+ page hand-out of lecture notes for the biochemistry class has co-authored a manual for the Chemistry for Health Sciences Laboratory and is working on a manual for the Biochemistry laboratory. To increase availability to the students, she conducts study sessions and distributes her home telephone number. Some students have attributed some of success to hand-outs distributed, graded homework and oral presentation assignments. A Biomaterials course will be developed that introduces implantology and describes the methods for choosing and characterizing materials for specific biological applications. The PI has already developed several new laboratory exercises for her biochemistry laboratory course and will prepare additional exercises that combine science disciplines. ***