ABSTRACT
Braun's lipoprotein (Lpp) plays a major role in stabilizing the integrity of the cell envelope in Escherichia coli, as it provides a covalent cross-link between the outer membrane and the peptidoglycan layer. An important challenge in elucidating the physiological role of Lpp lies in attaining a detailed understanding of its distribution on the peptidoglycan layer. Here, using atomic force microscopy, we visualized Lpp directly on peptidoglycan sacculi. Lpp is homogeneously distributed over the outer surface of the sacculus at a high density. However, it is absent at the constriction site during cell division, revealing its role in the cell division process with Pal, another cell envelope-associated protein. Collectively, we have established a framework to elucidate the distribution of Lpp and other peptidoglycan-bound proteins via a direct imaging modality.
Subject(s)
Escherichia coli , Lipoproteins , Microscopy, Atomic Force , Molecular Imaging , Bacterial Outer Membrane Proteins/metabolism , Cell Membrane/metabolism , Escherichia coli/chemistry , Lipoproteins/chemistry , Peptidoglycan/chemistry , Molecular Imaging/methodsABSTRACT
A highly diastereo- and enantioselective [4 + 1] cycloaddition of para-quinone methides to α-halogenated ketones was realized by bifunctional phosphonium salt catalysis, furnishing functionalized 2,3-dihydrobenzofurans in high yields and excellent stereoselectivities (>20:1 dr and up to >99.9% ee). Mechanistic observations suggested that the reaction underwent a cascade intermolecular substitution/intramolecular 1,6-addition process. DFT calculations revealed the presence of multiple H-bonding interactions between the catalyst and the enolate intermediate in the stereodetermining transition states.
ABSTRACT
The first enantioselective aza-Darzens reaction of cyclic imines with α-halogenated ketones was realized under mild reaction conditions by using amino-acid-derived bifunctional phosphonium salts as phase-transfer promoters. A variety of structurally dense tri- and tetrasubstituted aziridine derivatives, containing benzofused heterocycles as well as spiro-structures, were readily synthesized in high yields with excellent diastereo- and enantioselectivities (up to >20:1 d.r. and >99.9 % ee). The highly functionalized aziridine products could be easily transformed into different classes of biologically active compounds.
ABSTRACT
Renal ischemia-reperfusion (I/R) injury ranks as the primary cause of acute renal injury with severe morbidity and mortality. Side population (SP) cells have recently drawn increasing attention due to their critical role in injury repair and regeneration. Unfortunately, the underlying mechanism involved in renal I/R remains poorly elucidated. Here, pronounced increases of stromal cell-derived factor-1 (SDF-1) and its receptor CXC chemokine receptor 4 (CXCR4) were substantiated in I/R kidneys from C57BL/6 mice subjected to clamp the bilateral renal pedicles to mimic renal ischemia. Similar up-regulation of them was also determined in SP cells upon simulated ischemia/reperfusion (SI/R). In contrast to non-SP cells, SP cells exhibited higher viability, apoptosis resistance, chemotaxis, and paracrine actions following SI/R treatment, and these were further enhanced after SDF-1 stimulation. Interestingly, blocking CXCR4 signaling with AMD3100 notably ameliorated the above effects. Mechanism analysis corroborated that SDF-1/CXCR4 further induced the expression of ATP-binding cassette transporter ABCG2, an essential element for SP-mediated kidney regeneration after renal I/R injury. Moreover, AMD3100 pretreatment strikingly attenuated ABCG2 elevation in SP cells. Additionally, sonic hedgehog (SHH)-Gli 1 signaling was involved in SDF-1/CXCR4-mediated ABCG2 expression. When SP cells pretreated with AMD3100 were intravenously injected into I/R mice, SP cell-mediated decreases in blood urea nitrogen, serum creatinine, and histological score of kidney were noticeably attenuated, indicating that blocking CXCR4 pathway mitigated the therapeutic function of SP cells in renal I/R injury. Together, this research suggests that SDF-1/CXCR4 axis might act, via Shh-Gli1-ABCG2 signaling, as a positive regulator of SP cell-based therapies for renal I/R by Shh-Gli 1-ABCG2 signaling.
Subject(s)
ATP Binding Cassette Transporter, Subfamily G, Member 2/metabolism , Chemokine CXCL12/metabolism , Hedgehog Proteins/metabolism , Kidney/metabolism , Kidney/pathology , Receptors, CXCR4/metabolism , Reperfusion Injury/metabolism , Zinc Finger Protein GLI1/metabolism , ATP Binding Cassette Transporter, Subfamily G, Member 2/genetics , Animals , Chemokine CXCL12/genetics , Hedgehog Proteins/genetics , Male , Mice , Mice, Inbred C57BL , Receptors, CXCR4/genetics , Reperfusion Injury/genetics , Signal Transduction/genetics , Signal Transduction/physiology , Zinc Finger Protein GLI1/geneticsABSTRACT
This paper presents a kind of thermoresponsive polymeric magnetic particles for protein separations. The magnetofluids were directly encapsulated in hollow particles constructed by self-assembly of rod-coil poly(ethylene glycol)-poly(N-isopropylacrylamide)/α-cyclodextrin (PEG-PNIPAM/α-CD) complexes. The resulting particles showed reversible protein absorption/desorption capacity because the reversible thermo-sensitivity of PNIPAM. Above the lower critical solution temperature (LCST) of PNIPAM, these particles showed high absorptive capacities and adsorption was done at lower temperature. The protein-laden particles are readily removed from the feed solution in a magnetic field.
