INFLUENCE OF AA SUBSTITUTIONS ON STABILITY OF TETRAMER-DIMER INTERFACE OF HBS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 曼宁教授在东北大学的实验室和我们的实验室之间的持续和富有成效的合作仍在进行中。这项合作努力的实验目标是通过质谱学对人类血红蛋白多肽链中自然发生的或基因工程产生的突变进行完整的表征。最近，我们的实验室与J·埃里克·罗素教授的实验室开始了一个项目，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中影响氧向周围组织运输效率的长距离通讯。我们目前正在研究人类血红蛋白四聚体-二聚体界面附近某些氨基酸残基的关系。目前正在使用质谱学的三步法来表征血红蛋白中构成多肽链的多肽链，这三步方法包括：(1)用电喷雾电离质谱仪分析多肽链的完整质量；(2)用特定的蛋白酶消化多肽链并通过MALDI-TOF/MS获得肽图；(3)用电喷雾电离离子陷阱质谱仪对含有突变的多肽进行测序。

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.