Gregoire C, Rosinski-Chupin I, Rabillon J, Alzari P M, David B, Dandeu J P
Unité d'Immuno-Allergie, Département de Physiopathologie, Institut Pasteur, 28 rue du Dr Roux, 75024 Paris Cedex 15, France.
J Biol Chem. 1996 Dec 20;271(51):32951-9. doi: 10.1074/jbc.271.51.32951.
The gene encoding the major horse allergen, designated Equus caballus allergen 1 (Equ c1), was cloned from total cDNA of sublingual salivary glands by reverse transcription-polymerase chain reaction using synthetic degenerate oligonucleotides deduced from N-terminal and internal peptide sequences of the glycosylated hair dandruff protein. A recombinant form of the protein, with a polyhistidine tail, was expressed in Escherichia coli and purified by immobilized metal affinity chromatography. The recombinant protein is able to induce a passive cutaneous anaphylaxis reaction in rat, and it behaves similarly to the native Equ c1 in several immunological tests with allergic patients' IgE antibodies, mouse monoclonal antibodies, or rabbit polyclonal IgG antibodies. Amino acid sequence identity of 49-51% with rodent urinary proteins from mice and rats suggests that Equ c1 is a new member of the lipocalin superfamily of hydrophobic ligand-binding proteins that includes several other major allergens. An RNA blot analysis demonstrates the expression of mRNA Equ c1 in liver and in sublingual and submaxillary salivary glands.
编码主要马过敏原（命名为马变应原1，Equ c1）的基因，通过逆转录-聚合酶链反应，使用从糖基化毛发皮屑蛋白的N端和内部肽序列推导的合成简并寡核苷酸，从舌下唾液腺的总cDNA中克隆得到。一种带有多组氨酸尾巴的重组形式的该蛋白，在大肠杆菌中表达，并通过固定化金属亲和层析进行纯化。该重组蛋白能够在大鼠中诱导被动皮肤过敏反应，并且在与过敏患者的IgE抗体、小鼠单克隆抗体或兔多克隆IgG抗体进行的多项免疫试验中，其表现与天然的Equ c1相似。与来自小鼠和大鼠的啮齿动物尿蛋白的氨基酸序列一致性为49%-51%，这表明Equ c1是亲脂素疏水配体结合蛋白超家族的一个新成员，该超家族还包括其他几种蛋白。RNA印迹分析表明，Equ c1的mRNA在肝脏以及舌下和颌下唾液腺中表达。
