ON STABILITY OF TETRAMER-DIMER INTERFACE OF HUMAN HEMOGLOBINS

人血红蛋白四聚体-二聚体界面的稳定性研究

• 批准号: 7355140
• 负责人: JAMES M MANNING
• 金额: $ 0.25万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7355140
• 关键词: STABILITY; TETRAMER; DIMER; INTERFACE; HUMAN

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. An ongoing and productive collaboration is still in progress between Prof. Manning?s laboratory at Northeastern University and ours. The experimental goal of this collaborative endeavor is at the complete characterization by mass spectrometry of either naturally occurring or genetically engineered mutations in the polypeptide chains of human hemoglobins. Recently, our laboratories began a project with Prof. J. Eric Russell?s laboratory to study embryonic hemoglobins produced in transgenic mice. Our studies aim at a better understanding of the long-range communications within hemoglobin tetramers which affect the efficiency of the oxygen transport to peripheral tissues. We are currently investigating the relationship of certain amino acid residues near the tetramer-dimer interface of human hemoglobin. Mass spectrometry is being used to characterize the constituting polypeptide chains in hemoglobin using a three-step approach that consists in (1) analyzing the intact mass of the polypeptide chains by electrospray ionization mass spectrometry; (2) digesting the polypeptide chains with a specific proteinase and obtaining peptide maps by MALDI-TOF/MS; and (3) sequencing the peptides containing the mutations by mass spectrometric fragmentation on an electrospray ionization ion trap mass spectrometer. The following manuscript has been submitted for publication: MANNING, L. R.; RUSSELL, J. E.; PADOVAN, J. C.; CHAIT, B. T.; MANNING, J. M. ?-subunits in human embryonic hemoglobin Portland-2 (?2?2) weaken all subunit interactions relative to tetramers having ? subunits in adult hemoglobin A (?2?2).

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。曼宁教授和其他研究人员之间仍在进行持续和富有成效的合作？东北大学的实验室和我们的实验室。这项合作奋进的实验目标是通过质谱法对人血红蛋白多肽链中天然存在的或基因工程改造的突变进行完整表征。最近，我们的实验室开始了一个项目与教授。该实验室研究转基因小鼠产生的胚胎血红蛋白。我们的研究旨在更好地了解血红蛋白四聚体内的长距离通信，这会影响氧气运输到外周组织的效率。我们目前正在研究人类血红蛋白的四聚体-二聚体界面附近的某些氨基酸残基的关系。质谱法被用于使用三步法表征血红蛋白中的组成多肽链，该三步法包括（1）通过电喷雾电离质谱法分析多肽链的完整质量;（2）用特异性蛋白酶消化多肽链并通过MALDI-TOF/MS获得肽图;和（3）在电喷雾电离离子阱质谱仪上通过质谱片段化对含有突变的肽进行测序。以下手稿已提交出版：曼宁，L。的R.;拉塞尔，J.E.;帕多万;柴特，B。T.;曼宁?-人胚胎血红蛋白波特兰-2（？）两个？2)削弱所有亚基相互作用相对于四聚体有？成人血红蛋白A亚基（？两个？2）。