TRACHEAL ANTIMICROBIAL PEPTIDE, A CYSTEINE-RICH PEPTIDE FROM MAMMALIAN TRACHEAL MUCOSA - PEPTIDE ISOLATION AND CLONING OF A CDNA
TRACHEAL ANTIMICROBIAL PEPTIDE, A CYSTEINE-RICH PEPTIDE FROM MAMMALIAN TRACHEAL MUCOSA - PEPTIDE ISOLATION AND CLONING OF A CDNA
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DOI：
10.1073/pnas.88.9.3952
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发表时间：
1991-05-01
影响因子：
11.1
通讯作者：
BEVINS, CL
中科院分区：
文献类型：
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作者：
DIAMOND, G;ZASLOFF, M;BEVINS, CL
牛气管粘膜提取物中含有丰富的抗菌肽。 的38个氨基酸的肽，我们命名为气管抗菌肽（TAP），分离的顺序使用的尺寸排阻，离子交换，和反相色谱分馏使用抗菌活性作为功能测定。 产率几乎等于2 μ g/g湿粘膜。 通过肽和cDNA分析的组合确定完整的肽序列。 TAP的氨基酸序列为H-Asn-Pro-Val-Ser-Cys-Val-Arg-Asn-Lys-Gly-Ile-Cys-Val-Pro-Ile-Arg-Cys-Pro-Gly-Ser-Met-Lys-Gln-Ile-Gly-Thr-Cys-Val-Gly-Arg-Ala-Val-Lys-Cys-Cys-Arg-Lys-Lys-OH。 分离的肽的质谱分析与该序列一致，并表明6个半胱氨酸残基参与分子内二硫键的形成。 大小，基本电荷，和三个分子内二硫键的存在是类似的，但明显不同于，防御素，一个良好的特点，从哺乳动物循环吞噬细胞的抗菌肽类。 预测推定的TAP前体相对较小（64个氨基酸），成熟肽位于羧基末端，并被短的推定前肽区域和框内终止密码子包围。 编码该肽的mRNA在呼吸道粘膜中比在整个肺组织中更丰富。 纯化的多肽对大肠杆菌、金黄色葡萄球菌、肺炎克雷伯菌和铜绿假单胞菌具有体外抗菌活性。 此外，该肽对白色念珠菌有活性，表明具有广谱活性。 基于结构和活性，该肽似乎是在动物、昆虫和植物中发现的富含半胱氨酸的阳离子抗微生物肽组的成员。 从哺乳动物呼吸道粘膜中分离TAP可能为我们了解这一重要组织的宿主防御提供了见解。
Extracts of the bovine tracheal mucosa have an abundant peptide with potent antimicrobial activity. The 38-amino acid peptide, which we have named tracheal antimicrobial peptide (TAP), was isolated by a sequential use of size-exclusion, ion-exchange, and reverse-phase chromatographic fractionations using antimicrobial activity as a functional assay. The yield was almost-equal-to 2-mu-g/g of wet mucosa. The complete peptide sequence was determined by a combination of peptide and cDNA analysis. The amino acid sequence of TAP is H-Asn-Pro-Val-Ser-Cys-Val-Arg-Asn-Lys-Gly-IIe-Cys-Val-Pro-Ile-Arg-Cys-Pro-Gly-Ser-Met-Lys-Gln-Ile-Gly-Thr-Cys-Val-Gly-Arg-Ala-Val-Lys-Cys-Cys-Arg-Lys-Lys-OH. Mass spectral analysis of the isolated peptide was consistent with this sequence and indicated the participation of six cysteine residues in the formation of intramolecular disulfide bonds. The size, basic charge, and presence of three intramolecular disulfide bonds is similar to, but clearly distinct from, the defensins, a well-characterized class of antimicrobial peptides from mammalian circulating phagocytic cells. The putative TAP precursor is predicted to be relatively small (64 amino acids), and the mature peptide resides at the extreme carboxyl terminus and is bracketed by a short putative propeptide region and an in-frame stop codon. The mRNA encoding this peptide is more abundant in the respiratory mucosa than in whole lung tissue. The purified peptide had antibacterial activity in vitro against Escherichia coli, Staphylococcus aureus, Klebsiella pneumonia, and Pseudomonas aeruginosa. In addition, the peptide was active against Candida albicans, indicating a broad spectrum of activity. This peptide appears to be, based on structure and activity, a member of a group of cysteine-rich, cationic, antimicrobial peptides found in animals, insects, and plants. The isolation of TAP from the mammalian respiratory mucosa may provide insight into our understanding of host defense of this vital tissue.