Kubrycht J, Borecký J, Sigler K
Center of Occupational Diseases, National Institute of Public Health, 100 42 Prague, Czechia.
Folia Microbiol (Praha). 2002;47(4):319-58. doi: 10.1007/BF02818689.
Forty original sequences of peptide substrates and inhibitors of protein kinases and phosphatases were aligned in a chain matrix without artificial gaps. Fifteen protein kinase peptide substrates and inhibitors (PKSI peptides) contained a common dipeptide ArgArg and also additional important tetra-, tri- and dipeptide homologies. Three further peptide substrates were significantly similar to these peptides but lacked the ArgArg dipeptide. Sequence comparison of individual PKSI peptides revealed probabilistically restricted consensus sequence--PKSI motif--comprising 8 homologous and 13 non-randomly distributed amino acids without considering mutation analysis. This template motif was compared with the consensus sequences of 12 different immunoglobulin domains. In 11 of 12 these domains, the starts of homologous segments were found at nearly the same domain related sites, beginning with serine. A single-triplet mutation of any of the first two triplet bases that encode equally localized amino acids in each of the two sequence sets (PKSI and Ig) revealed additional homologies with the other set. A primary derived motif version composed of 9 homologous and seven non-randomly distributed amino acids was consequently established by its feedback projection into the original sequence sets. This procedure yielded a second preliminary motif version (revised motif) formed by a sequence of 9 homologous amino acids and two non-randomly distributed amino acids. In addition, three shorter oligopeptide motifs called important stereotypes were derived, based on repeated homology between Ig chains and the revised motif. The most extensive similarities in terms of these stereotypes occurred in the CH2 and CH4 domains of Ig peptides, and inhibitors of cAMP dependent protein kinase and protein kinase A. Further comparisons based on a reference sequence set arranged with the aid of feedback projection revealed a lower similarity between variable Ig chains reflected in a decreased number of homologous amino acids. Two final motif versions, FMC and FMV, were found in two different subsets of constant and variable Ig chains, respectively. FMC was composed of seven homologous and one non-randomly distributed amino acids forming the dispersed structure STLR(C)LVSD, whereas 6 homologous and one questionable amino acid constituted FMV. Only CH4 and CH1 domain segments contained all five high-incidence amino acids, which represented a higher level of similarity than homologous amino acids of all preliminary and final motifs. Four such amino acids were present also in three PKSI peptides. All similarities described here occur in domain segments positionally overlapping with the CDR1 region of variable chains. The results are discussed in terms of immunoglobulin evolution, the position of Fc receptor binding sites and degeneration or mutability of the triplets of motif-constituting amino acids.
40个蛋白激酶和磷酸酶的肽底物及抑制剂的原始序列在没有人为空位的链矩阵中进行比对。15个蛋白激酶肽底物及抑制剂(PKSI肽)含有一个共同的二肽ArgArg,并且还有额外重要的四肽、三肽和二肽同源性。另外三个肽底物与这些肽显著相似,但缺少ArgArg二肽。对各个PKSI肽的序列比较揭示了概率受限的共有序列——PKSI基序——由8个同源且13个非随机分布的氨基酸组成,未考虑突变分析。将这个模板基序与12个不同免疫球蛋白结构域的共有序列进行比较。在12个结构域中的11个中,同源片段的起始在几乎相同的与结构域相关的位点,从丝氨酸开始。对编码两个序列集(PKSI和Ig)中定位相同氨基酸的前两个三联体碱基中的任何一个进行单三联体突变,揭示了与另一个序列集的额外同源性。因此,通过将其反馈投影到原始序列集中,建立了一个由9个同源且7个非随机分布的氨基酸组成的一级衍生基序版本。这个过程产生了第二个初步基序版本(修订基序),由9个同源氨基酸和2个非随机分布的氨基酸序列组成。此外,基于Ig链与修订基序之间的重复同源性,衍生出了三个较短的寡肽基序,称为重要模式。在Ig肽的CH2和CH4结构域以及cAMP依赖性蛋白激酶和蛋白激酶A的抑制剂方面,这些模式的相似性最为广泛。基于借助反馈投影排列的参考序列集进行的进一步比较显示,可变Ig链之间的相似性较低,反映在同源氨基酸数量减少。在恒定和可变Ig链的两个不同子集中分别发现了两个最终基序版本,FMC和FMV。FMC由7个同源且1个非随机分布的氨基酸组成,形成分散结构STLR(C)LVSD,而FMV由6个同源和1个可疑氨基酸组成。只有CH4和CH1结构域片段包含所有五个高发生率氨基酸,这代表了比所有初步和最终基序的同源氨基酸更高的相似性水平。在三个PKSI肽中也存在四个这样的氨基酸。这里描述的所有相似性都出现在与可变链的CDR1区域位置重叠的结构域片段中。从免疫球蛋白进化、Fc受体结合位点的位置以及构成基序的氨基酸三联体的退化或可变性方面对结果进行了讨论。