非洲爪蟾两种卵化酶分子对卵黄膜作用机制的探讨

在分离纯化非洲爪蟾孵化酶时,得到了６０ＫＤ和４０ＫＤ两种分子,用卵化酶的特异性抗ＧＳＴ－ＵＶＳ．２抗体进行Ｗｅｓｔｅｒｎ杂交的结果证明二者均为孵化酶分子。６０ｋＤ分子很不稳定,在纯化过程中极易降解活性４０ＫＤ分子可能是由６０ＫＤ分子经过降解或自身降解丢失了两个ＣＵＢ重复区而形成的,而ＣＵＢ重复区很可能在对受精膜分子结构的修饰或改造中具有重要作用,在进一步验证其蛋白酶活性和生物活性时,发现二者几乎具     

非洲爪蟾孵化酶的cDNA结构及其部分生物化学特性的研究

以UVS．2为探针从第25期非洲爪蟾胚胎头背部的cDNA文库中筛选出了一个1．8kb的孵化酶基因(xhe),其转录产物最早出现于第17期胚胎的头背部,在第30期转录量达到高峰,随后便逐渐减少。该基因含有编码514个氨基酸的一个开框阅读框架,含有信号肽和原酶序列。所推测出的成熟蛋白有425个氨基酸,包括位于N一端的含有200个氨基酸的金属蛋白酶序列和位于C端的两个各110个氨基酸的CUB重复区。而UVS．2只代表该基因C端大约3／4的部分。同时还发现该酶分子量为60kDa,是一种胰蛋白酶类型的金属蛋白酶。它很不稳定,在纯化过程中极易降解为40kDa分子。60kDa分子具有很强的卵黄膜溶解活性和蛋白酶活性。其中CUB重复区很可能在介导卵黄膜和40kDa分子中起着重要作用,而40kDa分子很可能是在纯化操作过程中,由60kDa分子发生降解或自身降解丢失了两个CUB重复区而形成的,它只是60kDa分子中的一个金属蛋白酶主功能区,所以它没有卵黄膜溶解活性,尽管仍具有很强的蛋白酶活性。     

Decoding mechanisms by which silent codon changes influence protein biogenesis and function

ScopeSynonymous codon usage has been a focus of investigation since the discovery of the genetic code and its redundancy. The occurrences of synonymous codons vary between species and within genes of the same genome, known as codon usage bias. Today, bioinformatics and experimental data allow us to compose a global view of the mechanisms by which the redundancy of the genetic code contributes to the complexity of biological systems from affecting survival in prokaryotes, to fine tuning the structure and function of proteins in higher eukaryotes. Studies analyzing the consequences of synonymous codon changes in different organisms have revealed that they impact nucleic acid stability, protein levels, structure and function without altering amino acid sequence. As such, synonymous mutations inevitably contribute to the pathogenesis of complex human diseases. Yet, fundamental questions remain unresolved regarding the impact of silent mutations in human disorders. In the present review we describe developments in this area concentrating on mechanisms by which synonymous mutations may affect protein function and human health.PurposeThis synopsis illustrates the significance of synonymous mutations in disease pathogenesis. We review the different steps of gene expression affected by silent mutations, and assess the benefits and possible harmful effects of codon optimization applied in the development of therapeutic biologics.Physiological and medical relevanceUnderstanding mechanisms by which synonymous mutations contribute to complex diseases such as cancer, neurodegeneration and genetic disorders, including the limitations of codon-optimized biologics, provides insight concerning interpretation of silent variants and future molecular therapies.     

Comparative genome analysis of six malarial parasites using codon usage bias based tools

Codon usage bias (CUB) is an omnipresent phenomenon, which occurs in nearly all organisms. Previous studies of codon bias in Plasmodium species were based on a limited dataset. This study uses whole genome datasets for comparative genome analysis of six Plasmodium species using CUB and other related methods for the first time. Codon usage bias, compositional variation in translated amino acid frequency, effective number of codons and optimal codons are analyzed for P.falciparum, P.vivax, P.knowlesi, P.berghei, P.chabaudii and P.yoelli. A plot of effective number of codons versus GC3 shows their differential codon usage pattern arises due to a combination of mutational and translational selection pressure. The increased relative usage of adenine and thymine ending optimal codons in highly expressed genes of P.falciparum is the result of higher composition biased pressure, and usage of guanine and cytosine bases at third codon position can be explained by translational selection pressure acting on them. While higher usage of adenine and thymine bases at third codon position in optimal codons of P.vivax highlights the role of translational selection pressure apart from composition biased mutation pressure in shaping their codon usage pattern. The frequency of those amino acids that are encoded by AT ending codons are significantly high in P.falciparum due to action of high composition biased mutational pressure compared with other Plasmodium species. The CUB variation in the three rodent parasites, P.berghei, P.chabaudii and P.yoelli is strikingly similar to that of P.falciparum. The simian and human malarial parasite, P.knowlesi shows a variation in codon usage bias similar to P.vivax but on closer study there are differences confirmed by the method of Principal Component Analysis (PCA).

Abbreviations

CDS - Coding sequences, GC1 - GC composition at first site of codon, GC2 - GC composition at second site of codon, GC3 - GC composition at third site of codon, Ala - Alanine, Arg - Arginine, Asn - Asparagine, Asp - Aspartic acid, Cys - Cysteine, Gln - Glutamine Glu - Glutamic acid Gly - Glycine His - Histidine Ile - Isoleucine Leu - Leucine Lys - Lysine Met - Methionine Phe - Phenylalanine Pro - Proline Ser - Serine Thr - Threonine Trp - Tryptophan Tyr - Tyrosine Val - Valine.