SUPPORT FOR A GAS-PHASE PROTEIN/PEPTIDE SEQUENCER

支持气相蛋白质/肽测序仪

• 批准号: 3519385
• 负责人: Mark O Olson
• 金额: $ 15.5万
• 依托单位: UNIVERSITY OF MISSISSIPPI MED CTR
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-02-01 至 1987-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3519385
• 关键词: aminoacid analyzer; high performance liquid chromatography

We propose to establish a facility for micro-sequencing of proteins and peptides utilizing a gas-phase sequencer and a state-of-the-art high performance liquid chromatograph. At least eight investigators will use this facility for a variety of projects: 1) The primary structures and sites of phosphorylation and methylation of nucleolar nonhistone proteins C23, B23 and 160K will be analyzed. The facility will also be used in studying regions of interaction of these proteins with other macromolecules. 2) Peptides from the carboxyl-terminal region of eukaryotic polypeptide chain initiation factors eIF-2(Alpha) and eIF-4A will be sequenced. 3) Sequence information will be obtained on a 78 kDa poly(A) binding protein and the location of methylated lysine residues will be determined on elongation factor Tu from murine erythroleukemia cells. 4) Portions of amino-acyl-tRNA synthetases from the silkworm B. mori will be sequenced and active site regions will be identified after affinity labeling. 5) The facility will be used to assess the structural homogeneity of the Mu and L chains of mouse monoclonal antibodies. 6) To determine whether separate species of enzyme exist for each metal cofactor, sequence data will be obtained on manganese and iron superoxide dismutases from S. mutans. 7) Domains from catfish immunoglobulins will be sequenced to determine the structural basis for the distinct H and L chain subclasses. In projects 1 through 4 obtaining accurate sequence information is essential for cloning and DNA sequencing projects. In all projects the amounts of obtainable protein are limited, thereby creating the necessity for a high sensitivity protein sequencing facility.

我们建议建立一个蛋白质微测序设施 肽利用气相测序仪和最先进的高通量测序仪 高效液相色谱仪。 至少八名调查员将使用 该设施适用于各种项目：1）主要结构和 核仁非组蛋白的磷酸化和甲基化位点 将分析 C23、B23 和 160K。 该设施还将用于 研究这些蛋白质与其他蛋白质相互作用的区域 大分子。 2) 来自羧基末端区域的肽 真核多肽链起始因子 eIF-2(Alpha) 和 eIF-4A 将被测序。 3) 序列信息将在 78 kDa 上获得 Poly(A) 结合蛋白和甲基化赖氨酸残基的位置将 可以根据鼠红白血病细胞的延伸因子Tu进行测定。 4) 来自蚕 B. mori 的部分氨基-酰基-tRNA 合成酶将 进行测序，并在亲和后鉴定活性位点区域 标签。 5) 该设施将用于评估结构 小鼠单克隆抗体 Mu 和 L 链的同质性。 6) 至 确定每种金属辅因子是否存在不同种类的酶， 将获得锰和铁超氧化物歧化酶的序列数据 来自变形链球菌。 7) 对鲶鱼免疫球蛋白的结构域进行测序 确定不同 H 链和 L 链的结构基础 子类。 在项目 1 到 4 中获得准确的序列 信息对于克隆和 DNA 测序项目至关重要。 总共 预计可获得的蛋白质数量是有限的，从而创造了 高灵敏度蛋白质测序设备的必要性。

项目成果

RNA binding fragments from nucleolin contain the ribonucleoprotein consensus sequence.

• DOI: 10.1016/s0021-9258(18)60904-4
• 发表时间: 1987-08
• 期刊: The Journal of biological chemistry
• 作者: B. Bugler;H. Bourbon;B. Lapeyre;M. O. Wallace;J. Chang;F. Amalric;M. Olson

Characterization of a 48-kDa nucleic-acid-binding fragment of nucleolin.

核仁素 48 kDa 核酸结合片段的表征。

• DOI: 10.1111/j.1432-1033.1989.tb14581.x
• 发表时间: 1989
• 期刊: European journal of biochemistry
• 作者: Sapp,M;Richter,A;Weisshart,K;Caizergues-Ferrer,M;Amalric,F;Wallace,MO;Kirstein,MN;Olson,MO
• 通讯作者: Olson,MO

Ligand binding by murine IgM antibodies: intramolecular heterogeneity exists in certain, but not all, cases.

鼠 IgM 抗体的配体结合：在某些但并非全部情况下存在分子内异质性。

• DOI: 10.1016/0161-5890(88)90094-6
• 发表时间: 1988
• 期刊: Molecular immunology
• 影响因子: 3.6
• 作者: Pascual,D;Clem,LW
• 通讯作者: Clem,LW