Amino acid sequence of the T4 DNA helix-destabilizing protein.
Amino acid sequence of the T4 DNA helix-destabilizing protein.
复制标题
T4 DNA 螺旋不稳定蛋白的氨基酸序列。
DOI：
10.1073/pnas.77.8.4614
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发表时间：
1980
影响因子：
11.1
通讯作者：
Konigsberg,WH
中科院分区：
文献类型：
--
作者：
Williams,KR;LoPresti,MB;Setoguchi,M;Konigsberg,WH
由基因32编码的T4单链DNA结合蛋白的一级结构已通过对源自部分蛋白水解、溴化氰切割和用胰蛋白酶、胰凝乳蛋白酶和葡萄球菌蛋白酶消化的肽的手动和自动测序来确定。胰蛋白酶消化柠康酰化或琥珀酰化的基因32蛋白产生含有4、27、42、65和163个残基的五种肽，其可以通过Sephadex色谱分离。对每种胰蛋白酶肽进行自动测序，如果需要，进行更广泛的切割。完整的蛋白质含有301个氨基酸，分子量为33，487，并且可以通过有限的胰蛋白酶消化在赖氨酸21和253处特异性切割。先前的研究表明，具有+4电荷的"B"区（残基1 - 21）对于参与基因32蛋白与单链DNA的自缔合和协同结合的蛋白质-蛋白质相互作用是重要的。"A"区（残基254 - 301）与控制基因32蛋白的螺旋去稳定"活性"和与其他T4 DNA复制蛋白相互作用有关。A区的电荷为-10，此外，还含有两段由甘氨酸284分隔的不寻常的四个丝氨酸残基。位置73和115之间的区域含有75%的酪氨酸残基，并且可能对DNA结合很重要。
The primary structure of the T4 single-stranded DNA-binding protein coded by gene 32 has been determined by manual and autoated sequencing of peptides derived from partial proteolysis, cyanogen bromide cleavage, and digestion with trypsin, chymotrypsin, and staphylococcal protease. Tryptic digestion of citraconylated or succinylated gene 32 protein yields five peptides containing 4, 27, 42, 65, and 163 residues, which can be separated by Sephadex chromatography. Each of the tryptic peptides was subjected to automated sequencing and, if necessary, more extensive cleavage. The intact protein contains 301 amino acids, has a molecular weight of 33,487, and can be specifically cleaved at lysines 21 and 253 by limited trypsin digestion. Previous studies have shown that the "B" region (residues 1-21), which has a charge of +4, is important for the protein-protein interactions involved in gene 32 protein self-association and cooperaive binding to single-stranded DNA. The "A" region (residues 254-301) has been implicated in controlling the helix-destabilizing "activity" of gene 32 protein and in interacting with other T4 DNA replication proteins. The A region has a charge of -10 and, in addition, contains two unusual stretches of four serine residues separated by glycine 284. The region between positions 73 and 115 contains 75% of the tyrosine residues and may be important for DNA binding.