ANTI-PEPTIDE ANTIBODIES AND OLIGONUCLEOTIDE PROBES RECOGNIZING MAST CELL PROTEASE

基本上带电荷的丝氨酸蛋白酶和羧肽酶， 在神经pH下具有酶活性是分泌的主要成分， 人肥大细胞的颗粒。 虽然只有两种丝氨酸蛋白酶具有这种功能， 我们最近报道， 小鼠肥大细胞的分泌颗粒含有AT 至少六种丝氨酸蛋白酶。 这些小鼠肥大细胞蛋白酶来源于 但是它们的肽序列中有一些区域 100%相同 后一项发现引发了人们的担忧， 使用26-32 kDa特异性人肥大细胞丝氨酸产生的抗体 蛋白酶将与同源但不同的丝氨酸蛋白酶交叉反应 因为被该抗体识别的表位位于保守的 蛋白酶的区域。 DNA和抗肽抗体可以识别 不同的人肥大细胞蛋白及其相应的转录物， 关键试剂，以解决一些问题， mRNA含量与翻译和翻译后的关系 这些免疫细胞中的特定蛋白质的修饰。 照经上所 在这项研究项目中提出，使抗肽抗体和基因- 识别人羧肽酶A的特异性寡核苷酸探针， 不同的丝氨酸蛋白酶，存在于分泌颗粒中， 人肥大细胞 然后将这些试剂用于人肥大细胞的表型分析。 在身体过敏，系统性肥大细胞增多症， 和大疱性类天疱疮，并研究这些疾病的各种生化方面， 在培养的人类肥大细胞中的酶。

Basically charged, serine proteases and carboxypeptidases that are enzymatically active at neural pH are major constituents of the secretory granules of human mast cells. Although only two serine proteases have so far been identified in human mast cells, we recently reported that the secretory granules of mouse mast cells contain various combinations of at least six serine proteases. These mouse mast cell proteases are derived from distinct genes, but they have regions within their peptide sequences that are 100% identical. This latter finding raised concerns that antibodies produced using a 26-32 kDa specific human mast cell serine protease will cross react against a homologous but distinct serine protease because an epitope recognized by that antibody resides in the conserved region of the protease. DNAs and anti-peptide antibodies that recognize different human mast cell proteins and their corresponding transcripts are critical reagents for addressing a number of questions regarding the relationship of mRNA content to translation and post-translational modification of specific proteins in these immune cells. Thus, it is proposed in this research project to make anti-peptide antibodies and gene- specific oligonucleotide probes that recognize human carboxypeptidase A and distinct serine proteases that are present in the secretory granules of human mast cells. These reagents will then be used to phenotype human mast cells in vivo in patients with physical allergies, systemic mastocytosis, and bullous pemphigoid, and to study various biochemical aspects of these enzymes in cultured human mast cells.