A novel human airway mucin cDNA encodes a protein with unique tandem-repeat organization.
A novel human airway mucin cDNA encodes a protein with unique tandem-repeat organization.
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一种新型人气道粘蛋白 cDNA 编码具有独特串联重复组织的蛋白质。
DOI：
10.1042/bj3000295
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Sachdev,GP
中科院分区：
文献类型：
--
作者：
Shankar,V;Gilmore,MS;Elkins,RC;Sachdev,GP
采用高特异性亲和纯化的抗去糖基化人气管支气管粘蛋白的多克隆抗体，从正常人气管mRNA制备的Uni-ZAP cDNA表达文库中选择免疫反应克隆。三个阳性克隆中最大的一个被命名为pAM1，它与多克隆抗体反应强烈，并被进一步表征。序列分析显示，该cDNA全长941 bp，编码313个氨基酸的多肽。碱基3-892由不完善的41个核苷酸串联重复序列（CCAGGAGGGGACACCGGGTTCACGAGCTGCCCACGCCCTCT）组成，编码一种独特的多肽，具有两种类型的一致重复序列，TSCPRPLQEGTRV和TSCPRPLQEGTPGSRAAHALSRRGHRVHELPTSSPGGDTGF。推导出的氨基酸序列的总体组成与粘蛋白核心相符，富含丝氨酸、苏氨酸、脯氨酸、甘氨酸和丙氨酸（约51%）。用粘蛋白cDNA探针的Northern blots与正常人气管和囊性纤维化支气管分离的RNA显示出强烈的多分散杂交带。这些数据表明，由克隆pAM1编码的粘蛋白代表了一种独特的肽组织类型，这在迄今为止报道的粘蛋白cdna中尚未被描述。
Highly specific affinity-purified polyclonal antibodies against deglycosylated human tracheobronchial mucin was used to select immunoreactive clones from a Uni-ZAP cDNA expression library prepared from normal human tracheal mRNA. The largest of three positive clones, designated pAM1, which reacted strongly with the polyclonal antibodies, was further characterized. Sequence analyses revealed a partial 941 bp cDNA that encoded a 313-amino-acid polypeptide. Bases 3-892 consisted of imperfect 41-nucleotide tandem repeats (CCAGGAGGGGACACCGGGTTCACGAGCTGCCCACGCCCTCT) that encoded a unique polypeptide with two types of consensus repeats, TSCPRPLQEGTRV and TSCPRPLQEGTPGSRAAHALSRRGHRVHELPTSSPGGDTGF. The overall composition of the deduced amino acid sequence matched that expected for a mucin protein core and is rich in serine, threonine, proline, glycine and alanine (approximately 51%). Northern blots probed with the mucin cDNA exhibited intense polydisperse hybridization bands with RNA isolated from normal human trachea and cystic-fibrosis bronchus. The data indicate that mucin encoded by clone pAM1 represents a unique type of peptide organization which has not been described in mucin cDNAs reported thus far.