HPTLC coupled to ESI-Tandem MS for identifying phospholipids associated to membrane proteins in photosynthetic purple bacteria

Lapieza, M.P. ; Jungas, C. ; Savirón, M. ; Jarne, C. ; Membrado, L. ; Vela, J. (Universidad de Zaragoza) ; Orduna, J. (Universidad de Zaragoza) ; Garriga, R. (Universidad de Zaragoza) ; Galbán, J. (Universidad de Zaragoza) ; Cebolla, V.L.

Resumen: High-performance thin-layer chromatography (HPTLC)-densitometry was directly combined with electrospray (ESI) tandem mass spectrometry for obtaining rapid and relevant structural identification of phospholipids (PL) species associated to membrane proteins (MP), in non-sulfur, purple bacteria having photosynthetic activity. Thus, species belonging to phosphatidylcholines (PC), phosphatidylethanolamines (PE), cardiolipins (CL) and phosphatidylglycerols (PG) associated to MP were investigated in bacterial membrane extracts from Rhodobacter (Rb.) blasticus, Rhodospirillum (R.) rubrum and Rhodobaca (Rbc.) bogoriensis, as well as those which are bound to a purified MP-photosynthetic complex from Rbc. bogoriensis.   PL-classes were separated using a 7-step gradient-solvent sequence with a previous acid plate preconditioning, using Automated Multiple Development. Band zones of the plate corresponding to PL classes were selected to ensure their direct transfer to ion-trap MS equipment through an elution-based interface.   Under the studied conditions, ESI + -MS spectra of PC and CL mostly showed sodium adducts ([M + Na] + ) and [M-2H + 3Na] + , respectively, when recorded from the plate. The respective sodium adducts were fragmented in the ion-trap, and sodium remained as the charge of the fragment ions, thus being useful for their structural identification through MS/MS. ESI - -MS and MS/MS spectra of CL were also obtained as [M-2H] 2- , as well as those of PE and PG species as [M-H] - and [M] - , respectively.   In this way, relative composition profiles of each studied PL-class by ESI-MS, and further identification of individual PL and the molecular species belonging to each of them by MS/MS were obtained.

Idioma: Inglés

DOI: 10.1080/10826076.2018.1561465

Año: 2019

Publicado en: JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 42, 1-2 (2019), [9 pp]
ISSN: 1082-6076

Factor impacto JCR: 0.992 (2019)

Categ. JCR: CHEMISTRY, ANALYTICAL rank: 75 / 86 = 0.872 (2019) - Q4 - T3
Categ. JCR: BIOCHEMICAL RESEARCH METHODS rank: 74 / 77 = 0.961 (2019) - Q4 - T3

Factor impacto SCIMAGO: 0.268 - Pharmaceutical Science (Q2) - Analytical Chemistry (Q3) - Clinical Biochemistry (Q4) - Biochemistry (Q4)

Financiación: info:eu-repo/grantAgreement/ES/DGA-FSE/E25-17R

Tipo y forma: Article (Published version)

Área (Departamento): Área Química Analítica (Dpto. Química Analítica)
Área (Departamento): Área Química Orgánica (Dpto. Química Orgánica)
Área (Departamento): Área Química Física (Dpto. Química Física)