To monitor structural changes during the binding of Pi to the active site of mammalian alkaline phosphatase in water medium, reaction-induced infrared spectroscopy was used. The interaction of Pi with alkaline phosphatase was triggered by a photorelease of ATP from the inactive P(3)-[1-(2-nitrophenyl)]ethyl ester of ATP. After photorelease, ATP was sequentially hydrolyzed by alkaline phosphatase giving rise to adenosine and three Pi. Although a phosphodiesterase activity was detected prior the photorelease of ATP, it was possible to monitor the structural effects induced by Pi binding to alkaline phosphatase. Interactions of Pi with alkaline phosphatase were evidenced by weak infrared changes around 1631 and at 1639 cm(-1), suggesting a small distortion of peptide carbonyl backbone. This result indicates that the motion required for the formation of the enzyme-phosphate complex is minimal on the part of alkaline phosphatase, consistent with alkaline phosphatase being an almost perfect enzyme. Photoproduct 2-nitrosoacetophenone may bind to alkaline phosphatase in a site other than the active site of bovine intestinal alkaline phosphatase and than the uncompetitive binding site of L-Phe in bovine intestinal alkaline phosphatase, affecting one-two amino acid residues.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1304289 | PMC |
| http://dx.doi.org/10.1529/biophysj.103.034116 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Mimicking Nature: Effect of Architectural Design Inspired by Cancellous Bone on the Biological Response of hMSC Cultured on Titanium Scaffolds Fabricated by Laser Beam Powder Bed Fusion.
J Biomed Mater Res A
January 2025
Faculty of Materials Science and Engineering, Warsaw University of Technology, Warsaw, Poland.
Bone tissue regeneration can be affected by various architectonical features of 3D porous scaffold, for example, pore size and shape, strut size, curvature, or porosity. However, the design of additively manufactured structures studied so far was based on uniform geometrical figures and unit cell structures, which often do not resemble the natural architecture of cancellous bone. Therefore, the aim of this study was to investigate the effect of architectonical features of additively manufactured (aka 3D printed) titanium scaffolds designed based on microtomographic scans of fragments of human femurs of individuals of different ages on in vitro response of human bone-derived mesenchymal stem cells (hMSC).
View Article and Find Full Text PDFLithium-doped calcium silicate cement regulates the immune microenvironment and promotes M2 macrophage polarization for enhancing bone regeneration.
J Biol Eng
January 2025
Department of Biomedical Engineering, China Medical University, Taichung, 406040, Taiwan.
Bone defects present a significant challenge in orthopedics and trauma surgery, necessitating innovative approaches to stimulate effective bone regeneration. This study investigated the potential of lithium-doped calcium silicate (LiCS) cement to enhance bone regeneration and modulate the immune microenvironment to promote tissue repair. We synthesized a LiCS ceramic powder and performed comprehensive analyses of its physicochemical properties, including phase composition, morphology, setting time, and mechanical strength.
View Article and Find Full Text PDFFAecal micRobiota transplantation in primary sclerosinG chOlangitis (FARGO): study protocol for a randomised, multicentre, phase IIa, placebo-controlled trial.
BMJ Open
January 2025
National Institute of Health and Care Research (NIHR) Birmingham Biomedical Research Centre (BRC) Center for Liver and Gastrointestinal Research, University of Birmingham, Birmingham, England, UK
Introduction: Primary sclerosing cholangitis (PSC) is the classical hepatobiliary manifestation of inflammatory bowel disease (IBD). The strong association between gut and liver inflammation has driven several pathogenic hypotheses to which the intestinal microbiome is proposed to contribute. Pilot studies of faecal microbiota transplantation (FMT) in PSC and IBD are demonstrated to be safe and associated with increased gut bacterial diversity.
View Article and Find Full Text PDFComparison of Clinical and Pathologic Features of Antimitochondrial Antibodies-negative Primary Biliary Cirrhosis and Cholestatic Type Drug-induced Liver Injury.
J Clin Gastroenterol
December 2024
Department of Infectious Diseases, The First Affiliated Hospital of Nanchang University, Nanchang, China.
Aim: To compare the respective clinical and pathologic features of antimitochondrial antibodies-negative (AMA-negative) primary biliary cirrhosis (PBC) and cholestatic type drug-induced liver injury (DILI) for clinical differential diagnosis.
Patients And Methods: Clinical data from 23 patients with AMA-negative PBC and 39 patients with cholestatic type DILI, treated at our hospital between January 2013 and January 2024, were collected and retrospectively analyzed.
Results: The cholestatic type DILI group exhibited a higher incidence of malaise and abdominal pain compared with the AMA-negative PBC group.
Comparison of the Effects of H37Rv and Infiltration on Tumor Necrosis Factor-Alpha and Interleukin-1 Beta Expression in Osteoblasts.
Surg Infect (Larchmt)
January 2025
First Clinical Medical College, Ningxia Medical University, Yinchuan, China.
This study aims to compare the expression levels of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) in osteoblasts infiltrated with H37Rv (H37Rv) and to understand the differential bone destruction in spinal tuberculosis (STB) versus spondylitis (BS). Primary osteoblasts were isolated and cultured from the cranial bones of 2-5 days old mice and characterized by alkaline phosphatase (ALP) staining and alizarin red staining (ARS). H37Rv and were cultured to the logarithmic phase, and transfection solutions were prepared.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!