RESUMEN
Elimination of autoreactive developing B cells is an important mechanism to prevent autoantibody production. However, how B cell receptor (BCR) signaling triggers apoptosis of immature B cells remains poorly understood. We show that BCR stimulation up-regulates the expression of the lysosomal-associated transmembrane protein 5 (LAPTM5), which in turn triggers apoptosis of immature B cells through two pathways. LAPTM5 causes BCR internalization, resulting in decreased phosphorylation of SYK and ERK. In addition, LAPTM5 targets the E3 ubiquitin ligase WWP2 for lysosomal degradation, resulting in the accumulation of its substrate PTEN. Elevated PTEN levels suppress AKT phosphorylation, leading to increased FOXO1 expression and up-regulation of the cell cycle inhibitor p27Kip1 and the proapoptotic molecule BIM. In vivo, LAPTM5 is involved in the elimination of autoreactive B cells and its deficiency exacerbates autoantibody production. Our results reveal a previously unidentified mechanism that contributes to immature B cell apoptosis and B cell tolerance.
Asunto(s)
Apoptosis , Tolerancia Inmunológica , Proteínas de la Membrana , Células Precursoras de Linfocitos B , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Proteína Forkhead Box O1/metabolismo , Humanos , Lisosomas/metabolismo , Proteínas de la Membrana/genética , Fosfohidrolasa PTEN/metabolismo , Células Precursoras de Linfocitos B/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ubiquitina-Proteína Ligasas/metabolismoRESUMEN
We present the finding of a dimeric ACE2 peptide mimetic designed through side chain cross-linking and covalent dimerization. It has a binding affinity of 16 nM for the SARS-CoV-2 spike RBD, and effectively inhibits the SARS-CoV-2 pseudovirus in Huh7-hACE2 cells with an IC50 of 190 nM and neutralizes the authentic SARS-CoV-2 in Caco2 cells with an IC50 of 2.4 µM. Our study should provide a new insight for the optimization of peptide-based anti-SARS-CoV-2 inhibitors.
Asunto(s)
Antivirales/farmacología , Fragmentos de Péptidos/farmacología , Peptidomiméticos/farmacología , SARS-CoV-2/efectos de los fármacos , Secuencia de Aminoácidos , Enzima Convertidora de Angiotensina 2/química , Antivirales/síntesis química , Antivirales/metabolismo , Línea Celular Tumoral , Humanos , Pruebas de Sensibilidad Microbiana , Fragmentos de Péptidos/síntesis química , Fragmentos de Péptidos/metabolismo , Peptidomiméticos/síntesis química , Peptidomiméticos/metabolismo , Unión Proteica , Dominios Proteicos , Glicoproteína de la Espiga del Coronavirus/química , Glicoproteína de la Espiga del Coronavirus/metabolismoRESUMEN
A series of cyclic Arg-rich mitochondria-penetrating peptides were prepared with variation in the macrocycle size and the chirality of Arg residues. A cyclic heptapeptide was demonstrated to be an efficient mitochondria-specific delivery vector for delivering membrane impermeable peptides.
Asunto(s)
Membrana Celular/metabolismo , Péptidos de Penetración Celular/metabolismo , Mitocondrias/metabolismo , Membrana Celular/química , Supervivencia Celular , Péptidos de Penetración Celular/química , Ciclización , Células HeLa , Humanos , Mitocondrias/química , Conformación MolecularRESUMEN
A new robust strategy was reported for the epimerization-free synthesis of C-terminal Cys-containing peptide acids through mercaptoethanol-mediated hydrolysis of peptide thioesters prepared in situ from peptide hydrazides. This simple-to-operate and highly efficient method avoids the use of derivatization reagents for resin modification, thus providing a practical avenue for the preparation of C-terminal Cys-containing peptide acids.
