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禽平滑肌S100A11(钙粒蛋白,S100C)的分子克隆与表达

Molecular cloning and expression of avian smooth muscle S100A11 (calgizzarin, S100C).

作者信息

Schönekess B O, Walsh M P

机构信息

Smooth Muscle Research Group, University of Calgary, AB, Canada.

出版信息

Biochem Cell Biol. 1997;75(6):771-5.

PMID:9599666
Abstract

S100A11 (calgizzarin or S100C), a member of the S100 family of Ca(2+)-binding proteins, was first identified in chicken gizzard smooth muscle and subsequently detected in several mammalian species and tissues. We now report the full-length coding sequence of avian smooth muscle S100A11. The cloned nucleotide sequence is 515 bases in length, which includes in-frame start and stop codons and encodes a protein of 101 amino acids. The chicken S100A11 sequence differs from human S100A11 at 25 positions (9 conserved) and is four residues shorter (overall identity 72.4%, similarity 81%). The protein contains two EF hand and conserved hydrophobic residues involved in dimer formation. Cloned avian S100A11 expressed in Escherichia coli and purified by Ca(2+)-dependent hydrophobic interaction chromatography and ion-exchange chromatography was recognized by polyclonal antibodies raised against tissue-purified protein and, like tissue-purified S100A11, bound 45Ca2+ in a gel overlay assay.

摘要

S100A11(钙粒蛋白或S100C)是Ca(2+)结合蛋白S100家族的成员之一,最初在鸡砂囊平滑肌中被鉴定出来,随后在几种哺乳动物的物种和组织中也被检测到。我们现在报告禽平滑肌S100A11的全长编码序列。克隆的核苷酸序列长度为515个碱基,其中包括符合读框的起始和终止密码子,编码一个由101个氨基酸组成的蛋白质。鸡S100A11序列与人类S100A11在25个位置上不同(9个保守),并且短四个残基(总体一致性为72.4%,相似性为81%)。该蛋白质包含两个EF手结构以及参与二聚体形成的保守疏水残基。在大肠杆菌中表达并通过Ca(2+)依赖性疏水相互作用色谱法和离子交换色谱法纯化的克隆禽S100A11,能被针对组织纯化蛋白产生的多克隆抗体识别,并且与组织纯化的S100A11一样,在凝胶覆盖试验中能结合45Ca2+。

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