Wattiez R, Hermans C, Bernard A, Lesur O, Falmagne P
Department of Biological Chemistry, University of Mons-Hainaut, Mons, Belgium.
Electrophoresis. 1999 Jun;20(7):1634-45. doi: 10.1002/(SICI)1522-2683(19990601)20:7<1634::AID-ELPS1634>3.0.CO;2-J.
Although bronchoalveolar lavage has been used as a research and clinical tools for more than two decades to investigate the cellular and soluble components of the lower respiratory tract, its exact protein composition has never been established. In this context, proteins of human bronchoalveolar lavage fluids (BALF), obtained by washing the epithelial lining fluid of the lungs with phosphate-buffered saline, were analyzed by two-dimensional electrophoresis (2-DE) under denaturing and reducing conditions. To characterize the widest amount of proteins, an analytical map of human bronchoalveolar lavage fluid proteins has been created from a pool of BALF from various patients. The resulting map comprises 211 silver-stained spots in the range of pI 3.5-10 and molecular mass 5-100 kDa. We identified 182 spots by microsequence analysis and by matching with human blood plasma and the Macrophage Like Cell line reference 2-DE maps available from the SWISS-2DPAGE database. The human bronchoalveolar lavage fluid was found to contain 61 different proteins or isoforms thereof. Most of the proteins had low molecular masses (< 35 kDa) and rather acidic isoelectric points (pI; 4 < pI < 7). The proteins in the lavage either are produced locally or originate from plasma. Two unknown proteins were identified and are currently under investigation.
尽管支气管肺泡灌洗作为一种研究和临床工具已被使用了二十多年，用于研究下呼吸道的细胞和可溶性成分，但其确切的蛋白质组成尚未确定。在此背景下，通过用磷酸盐缓冲盐水冲洗肺上皮衬液获得的人支气管肺泡灌洗流体（BALF）中的蛋白质，在变性和还原条件下通过二维电泳（2-DE）进行了分析。为了表征最广泛的蛋白质，已从不同患者的BALF池中创建了人支气管肺泡灌洗流体蛋白质的分析图谱。所得图谱在pI 3.5-10和分子量5-100 kDa范围内包含211个银染斑点。我们通过微序列分析并与人类血浆以及可从SWISS-2DPAGE数据库获得的巨噬细胞样细胞系参考2-DE图谱进行匹配，鉴定了182个斑点。发现人支气管肺泡灌洗流体含有61种不同的蛋白质或其同工型。大多数蛋白质具有低分子量（<35 kDa）和相当酸性的等电点（pI；4 <pI <7）。灌洗中的蛋白质要么在局部产生，要么源自血浆。鉴定出两种未知蛋白质，目前正在研究中。
