XPlex: An Effective, Multiplex Cross-Linking Chemistry for Acidic Residues

Autor(es)

Fioramonte, Mariana ^[1] ; Ramos de Jesus, Hugo Cesar ^[1] ; Ramos Ferrari, Allan Jhonathan ^[1] ; Lima, Diogo Borges ^[2] ; Drekener, Roberta Lopes ^[1] ; Duarte Correia, Carlos Roque ^[1] ; Oliveira, Luciana Gonzaga ^[1] ; da Costa Neves-Ferreira, Ana Gisele ^[3] ; Carvalho, Paulo Costa ^[4] ; Gozzo, Fabio Cesar ^[1]

Afiliação do(s) autor(es)

^[1] Univ Estadual Campinas, Inst Chem, CP 6154, BR-13083970 Campinas, SP - Brazil

^[2] Inst Pasteu, CNRS USR 2000, Mass Spectrometry Biol Unit, Paris - France

^[3] Fiocruz MS, Oswaldo Cruz Inst, Lab Toxinol, Rio De Janeiro - Brazil

^[4] Fiocruz MS, Carlos Chagas Inst, Lab Prote & Prot Engn, Curitiba, Parana - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Analytical Chemistry; v. 90, n. 10, p. 6043-6050, MAY 15 2018.
• Citações Web of Science: 6

Resumo

Cross-linking/Mass spectrometry (XLMS) is a consolidated technique for structural characterization of proteins and protein complexes. Despite its success, the cross-linking chemistry currently used is mostly based on N-hydroxysuccinimide (NHS) esters, which react primarily with lysine residues. One way to expand the current applicability of XLMS into several new areas is to increase the number of cross-links obtainable for a target protein. We introduce a multiplex chemistry (denoted XPlex) that targets Asp, Glu, Lys, and Ser residues. XPlex can generate significantly more cross-links with reactions occurring at lower temperatures and enables targeting proteins that are not possible with NHS ester-based cross-linkers. We demonstrate the effectiveness of our approach in model proteins as well as a target Lys-poor protein, SalBIII. Identification of XPlex spectra requires a search engine capable of simultaneously considering multiple cross-linkers on the same run; to achieve this, we updated the SIM-XL search algorithm with a search mode tailored toward XPlex. In summary, we present a complete chemistry/computational solution for significantly increasing the number of possible distance constraints by mass spectrometry experiments, and thus, we are convinced that XPlex poses as a real complementary approach for structural proteomics studies. (AU)

• Processo FAPESP: 14/50249-8 - Green chemistry: sustainable synthetic methods employing benign solvents, safer reagents, and bio-renewable feedstock
• Beneficiário: Arlene Gonçalves Corrêa
• Modalidade de apoio: Auxílio à Pesquisa - Programa Centros de Pesquisa em Engenharia
• Processo FAPESP: 16/13195-2 - Modelagem da estrutura de proteínas e de complexos protéicos usando dados de espectrometria de massas
• Beneficiário: Allan Jhonathan Ramos Ferrari
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 14/17264-3 - Novas fronteiras em proteômica estrutural: caracterizando estruturas de proteínas e complexos proteicos por espectrometria de massas
• Beneficiário: Fabio Cesar Gozzo
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 12/10862-7 - Proteômica Estrutural por Ligação Cruzada Acoplada a Espectrometria de Massas: Desenvolvimentos, Estudos Fundamentais e Aplicações.
• Beneficiário: Mariana Fioramonte
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 14/12727-5 - Genome mining em Streptomyces
• Beneficiário: Luciana Gonzaga de Oliveira
• Modalidade de apoio: Auxílio à Pesquisa - Regular