Analysis of secondary structure in proteins by chemical cross-linking coupled to MS

作者：Fioramonte Mariana; dos Santos Aline Mara; McIlwain Sean; Noble William S; Franchini Kleber G; Gozzo Fabio C^*

来源：Proteomics, 2012, 12(17): 2746-2752.

DOI：10.1002/pmic.201200040

• Alpha helix
• Beta sheet
• Cross-linking
• MS
• Protein secondary structure
• Technology

Chemical cross-linking is an attractive technique for the study of the structure of protein complexes due to its low sample consumption and short analysis time. Furthermore, distance constraints obtained from the identification of cross-linked peptides by MS can be used to construct and validate protein models. If a sufficient number of distance constraints are obtained, then determining the secondary structure of a protein can allow inference of the protein's fold. In this work, we show how the distance constraints obtained from cross-linking experiments can identify secondary structures within the protein sequence. Molecular modeling of alpha helices and beta sheets reveals that each secondary structure presents different cross-linking possibilities due to the topological distances between reactive residues. Cross-linking experiments performed with amine reactive cross-linkers with model alpha helix containing proteins corroborated the molecular modeling predictions. The cross-linking patterns established here can be extended to other cross-linkers with known lengths for the determination of secondary structures in proteins.

• 出版日期2012-8