Chandra Klara Kharisma Bunga, Nuringtyas Tri Rini, Raharjo Tri Joko
Department of Biotechnology, Faculty of Graduate School, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia.
BioTechnologia (Pozn). 2024 Dec 19;105(4):325-335. doi: 10.5114/bta.2024.145254. eCollection 2024.
is a genus of red algae found mainly in Asia. Various species of are distributed throughout Indonesia's marine waters, especially in coastal areas, and are cultivated for research and economic purposes. mainly consists of polysaccharides and pigments, which have hindered protein identification. The unique protein expressions have never been analyzed using a proteomic approach, and no reports are available on spp. amino acid sequences in Central Java. Based on this background, we aimed to explore protein characterization using unique peptide sequence analysis via LC-HRMS. The usage of liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) has been growing in biomedical and environmental sciences, offering high accuracy in protein detection. We followed the LC-HRMS standard protocol with an optimized precipitation procedure. TCA/acetone precipitation was used for protein purification, after which the precipitate was subjected to protein digestion to obtain small peptide fractions. Protein analysis results are presented as protein concentrations, molecular models, and peptide sequences. This experiment identified four sequences derived from the Rhodophyta database: TKKILDK (845.5455 Da), TVKSLLTK (889.5717 Da), ILVKTLK (814.5761 Da), and TGcGRSKR (921.4683 Da). This study reveals peptide sequences for spp., showing similarities with other red algae species, along with the functions of the peptide sequences. Furthermore, amino acid models of secondary structures were provided to support our findings.
是一种主要在亚洲发现的红藻属。的各种物种分布在印度尼西亚的海域，特别是在沿海地区，并被用于研究和经济目的而进行养殖。主要由多糖和色素组成，这阻碍了蛋白质的鉴定。其独特的蛋白质表达从未使用蛋白质组学方法进行分析，并且在中爪哇也没有关于 spp. 氨基酸序列的报道。基于此背景，我们旨在通过液相色谱 - 高分辨率质谱（LC - HRMS）使用独特的肽序列分析来探索蛋白质特征。液相色谱与高分辨率质谱联用（LC - HRMS）在生物医学和环境科学中的应用一直在增加，在蛋白质检测方面具有很高的准确性。我们遵循了带有优化沉淀程序的LC - HRMS标准方案。使用三氯乙酸/丙酮沉淀法进行蛋白质纯化，之后将沉淀物进行蛋白质消化以获得小肽片段。蛋白质分析结果以蛋白质浓度、分子模型和肽序列呈现。该实验鉴定出了来自红藻数据库的四个序列：TKKILDK（845.5455 Da）、TVKSLLTK（889.5717 Da）、ILVKTLK（814.5761 Da）和TGcGRSKR（921.4683 Da）。本研究揭示了 spp. 的肽序列，显示出与其他红藻物种的相似性以及肽序列的功能。此外，还提供了二级结构的氨基酸模型以支持我们的发现。
