Membrane fluidity adjusts the insertion of the transacylase PlsX to regulate phospholipid biosynthesis in Gram-positive bacteria

Autor(es)

Sastre, Diego E. ^[1] ; Basso, Luis G. M. ^[2] ; Trastoy, Beatriz ^[3] ; Cifuente, Javier O. ^[3] ; Contreras, Xabier ^{[4, 5, 6]} ; Gueiros-Filho, Frederico ^[7] ; de Mendoza, Diego ^[8] ; Navarro, Marcos V. A. S. ^[1] ; Guerin, Marcelo E. ^{[4, 3]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Fis Sao Carlos, Grp Biofis Mol Sergio Mascarenhas, Sao Carlos, SP - Brazil

^[2] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Fis, Ribeirao Preto, SP - Brazil

^[3] CIC BioGUNE Technol Pk Bizkaia, Struct Biol Unit, Derio 48160, Vizcaya - Spain

^[4] Ikerbasque, Basque Fdn Sci, Bilbao 48013 - Spain

^[5] Univ Basque Country, CSIC, Inst Biofis, Barrio Sarriena S-N, Leioa 48940, Bizkaia - Spain

^[6] Univ Basque Country, Dept Bioquim, Leioa 48940, Bizkaia - Spain

^[7] Univ Sao Paulo, Inst Quim, Dept Bioquim, Sao Paulo, SP - Brazil

^[8] Inst Biol Mol & Celular Rosario IBR, Rosario, Santa Fe - Argentina

• Tipo de documento: Artigo Científico
• Fonte: Journal of Biological Chemistry; v. 295, n. 7, p. 2136-2147, FEB 14 2020.
• Citações Web of Science: 2

Resumo

PlsX plays a central role in the coordination of fatty acid and phospholipid biosynthesis in Gram-positive bacteria. PlsX is a peripheral membrane acyltransferase that catalyzes the conversion of acyl-ACP to acyl-phosphate, which is in turn utilized by the polytopic membrane acyltransferase PlsY on the pathway of bacterial phospholipid biosynthesis. We have recently studied the interaction between PlsX and membrane phospholipids in vivo and in vitro, and observed that membrane association is necessary for the efficient transfer of acyl-phosphate to PlsY. However, understanding the molecular basis of such a channeling mechanism remains a major challenge. Here, we disentangle the binding and insertion events of the enzyme to the membrane, and the subsequent catalysis. We show that PlsX membrane binding is a process mostly mediated by phospholipid charge, whereas fatty acid saturation and membrane fluidity remarkably influence the membrane insertion step. Strikingly, the PlsX(L254E) mutant, whose biological functionality was severely compromised in vivo but remains catalytically active in vitro, was able to superficially bind to phospholipid vesicles, nevertheless, it loses the insertion capacity, strongly supporting the importance of membrane insertion in acyl-phosphate delivery. We propose a mechanism in which membrane fluidity governs the insertion of PlsX and thus regulates the biosynthesis of phospholipids in Gram-positive bacteria. This model may be operational in other peripheral membrane proteins with an unprecedented impact in drug discovery/development strategies. (AU)

• Processo FAPESP: 16/05203-5 - Como bactérias coordenam a biogênese de membranas com o crescimento e a divisão celular?
• Beneficiário: Frederico José Gueiros Filho
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/21583-0 - Desenvolvimento de novos antibióticos contra cepas super-resitentes de Staphylococcus aureus: identificação e caracterização de inibidores da enzima Diadenilato Ciclase
• Beneficiário: Marcos Vicente de Albuquerque Salles Navarro
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 14/00206-0 - Estrutura e função dos peptídeos de fusão da glicoproteína spike do SARS-CoV
• Beneficiário: Luís Guilherme Mansor Basso
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/13411-1 - Estudo de aciltransferases necessárias para a síntese de fosfolipídeos de membrana de bactérias Gram-positivas: um alvo atraente para a descoberta de drogas anti-bacterianas
• Beneficiário: Diego Emiliano Sastre
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado