ELECTRON MICROSCOPY OF MACROMOLECULAR STRUCTURES

大分子结构的电子显微镜

• 批准号: 3268609
• 负责人: HENRY S SLAYTER
• 金额: $ 12.62万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-04-01 至 1991-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3268609
• 关键词: antibody formation; antibody specificity; blood proteins; chemical structure; complement; conformation; electron microscopy; fibrinogen; fibronectins; gel electrophoresis; gel filtration chromatography; glycoproteins; human tissue; immunoglobulins; laboratory mouse; laboratory rabbit; lyophilization; macromolecule; membrane activity; membrane proteins; mucins; muscle proteins; myosins; polymers; radioimmunoassay; radiotracer; tissue /cell preparation

The proposed research will improve methods for high-resolution EM of individual macromolecules, and will pursue applications of these to cogent biological problems. Coating by thin refractory metal films effectively contrasts individual macromolecular substructure. Due to recent improvements in electron optics, grain size, rather than instrumental factors, currently limits ultimate resolution. Thus, we intend to produce fine-grain coatings which will fully exploit available resolving power, particularly applying high-contrast darkfield imaging. We will explore use of mixtures of refractory metals (which should limit possibilities for crystallization and thus reduce crystallite size) and of low temperature specimen supports (which should favor film continuity). Effects of solvent systems and drying conditions will also be studied. These techniques will be applied to a number of macromolecular systems, including myosin, fibrinogen, epiglycanin, retinol-binding protein, antigen-antibody complexes, bronchial mucins, and hamster female protein. We plan applications of the electronmicroscopic mapping approach, in which macromolecules are complexed with readily-resolvable, site-specific protein markers, to a number of problems. Chemically-defined sites in the fibrinogen molecule, including the important region which binds factor XIII, staphylococcin and blood platelets, will be located by complexing with specifically-directed Fabs or with monoclonal antibodies. Positions of the 4 carbohydrate residues in this molecule will be determined by complexing with specific lectins. We will further characterize cardiac myosin in a combined physical-chemical and electron microscopic study, as well as assessing the effects of various physiological conditions on function and structure of myosin S1 region, and polymeric forms. Cardiac myosin will be mapped with specific Fabs directed against subregions of these molecules (S1, S2, light chains, S1 subdomains and hinge) in order to elucidate topography of the myosin head region. Positions of specific epitopes and carbohydrate residues along epiglycanin (a mouse mammary tumor cell-surface glycoprotein) will be mapped by a series of specific lectins, and by rabbit and monoclonal IgG and IgM. The latter will be used to determine the specificities of these antibodies for epiglycanin, and for the monospecific site, respectively. Certain other large glycoproteins will also be mapped.

提出的研究将改进高分辨率电磁成像的方法

项目成果

Electron microscopy of cardiac myosin: its shape and properties as determined by the regulatory light chain.

心肌肌球蛋白的电子显微镜：其形状和特性由调节轻链决定。

• DOI: 10.1007/bf01578433
• 发表时间: 1987
• 期刊: Journal of muscle research and cell motility
• 影响因子: 2.7
• 作者: Margossian,SS;Slayter,HS
• 通讯作者: Slayter,HS

Proteolytic susceptibility of both isolated and bound light chains from various myosins to myopathic hamster protease.

各种肌球蛋白的分离和结合轻链对肌病仓鼠蛋白酶的蛋白水解敏感性。

• 发表时间: 1984
• 期刊: The Journal of biological chemistry
• 作者: Margossian,SS;Chantler,PD;Sellers,JR;Malhotra,A;Stafford,WF;Slayter,HS
• 通讯作者: Slayter,HS

Control of filament length by the regulatory light chains in skeletal and cardiac myosins.

通过骨骼和心脏肌球蛋白中的调节轻链控制丝长度。

• 发表时间: 1987
• 期刊: The Journal of biological chemistry
• 作者: Margossian,SS;Huiatt,TW;Slayter,HS
• 通讯作者: Slayter,HS

The molecular structure of lubricating glycoprotein-I, the boundary lubricant for articular cartilage.

• DOI: 10.1016/s0021-9258(19)69297-5
• 发表时间: 1981-06
• 期刊: The Journal of biological chemistry
• 作者: David A. Swann;H S Slayter;Fred H. Silver

A conformational transition in gizzard heavy meromyosin involving the head-tail junction, resulting in changes in sedimentation coefficient, ATPase activity, and orientation of heads.

砂囊重粒肌球蛋白中涉及头尾连接的构象转变，导致沉降系数、ATP酶活性和头部方向的变化。

• 发表时间: 1985
• 期刊: The Journal of biological chemistry
• 作者: Suzuki,H;Stafford3rd,WF;Slayter,HS;Seidel,JC
• 通讯作者: Seidel,JC