ABSTRACT
BACKGROUND: Proteins from the LytR-CpsA-Psr family are found in almost all Gram-positive bacteria. Although LCP proteins have been studied in other pathogens, their functions in enterococci remain uncharacterized. The Psr protein from Enterococcus hirae, here renamed LcpA, previously associated with the regulation of the expression of the low-affinity PBP5 and ß-lactam resistance, has been characterized. RESULTS: LcpA protein of E. hirae ATCC 9790 has been produced and purified with and without its transmembrane helix. LcpA appears, through different methods, to be localized in the membrane, in agreement with in silico predictions. The interaction of LcpA with E. hirae cell wall indicates that LcpA binds enterococcal peptidoglycan, regardless of the presence of secondary cell wall polymers. Immunolocalization experiments showed that LcpA and PBP5 are localized at the division site of E. hirae. CONCLUSIONS: LcpA belongs to the LytR-CpsA-Psr family. Its topology, localization and binding to peptidoglycan support, together with previous observations on defective mutants, that LcpA plays a role related to the cell wall metabolism, probably acting as a phosphotransferase catalyzing the attachment of cell wall polymers to the peptidoglycan.
Subject(s)
Bacterial Proteins/metabolism , Carrier Proteins/metabolism , Enterococcus hirae/metabolism , Peptidoglycan/metabolism , Repressor Proteins/metabolism , Amino Acid Sequence , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Bacterial Proteins/isolation & purification , Base Sequence , Carrier Proteins/chemistry , Carrier Proteins/genetics , Carrier Proteins/isolation & purification , Cell Membrane/metabolism , Cell Wall/metabolism , Cloning, Molecular , DNA, Bacterial , Enterococcus hirae/cytology , Enterococcus hirae/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Genes, Bacterial , Phosphotransferases/metabolism , Protein Interaction Maps , Recombinant Proteins , Repressor Proteins/chemistry , Repressor Proteins/genetics , Repressor Proteins/isolation & purification , beta-Lactam ResistanceABSTRACT
In this study, the authors used the Fujifilm Prescale Pressure Measuring System to measure the contact pressure and distribution at the jaws of laparoscopic grasping forceps. This data was then correlated with measured pressures at the forceps handles to understand the relationship between the surgeon's actuating pressure and that on the organ being manipulated. The purpose of this study is to create a database of tactile information to provide guidelines in defining minimally invasive surgery (MIS). This is expected to be important as today's society continues to progress in the use of automation, IoT, AI and MIS. In order to achieve the above, the authors developed an experimental device consisting of an actuator, a load cell and an MCU to stably actuate and control the handle side of grasping forceps. Target organs were simulated using triangular prisms of various silicone rubber materials. The experimental method involved actuating the handle side with preset pressure values for fixed time periods and using sensitive film to measure the pressure at the forceps tip. The film data was then scanned, processed and analyzed.