This protocol describes how to prepare rat liver rough microsomes that contain undegraded membrane-bound polysomes and can function very well in an in vitro translation system. It uses endogenous ribonuclease inhibitor in all steps, avoiding pelleting rough microsomes in all steps and sacrificing good recovery.
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http://dx.doi.org/10.1101/pdb.prot079970 | DOI Listing |
Arch Biochem Biophys
September 2024
Department of Physiology, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran. Electronic address:
Objectives: Potassium channels in the endoplasmic reticulum (ER) are crucial for maintaining calcium balance during calcium fluxes. Disruption in ER calcium balance leads to ER stress, implicated in diseases like diabetes and Alzheimer's disease (AD). However, limited data exists on ER potassium channels in excitable tissues such as the brain.
View Article and Find Full Text PDFJ Trace Elem Med Biol
December 2021
Department of Physics, Università Degli Studi di Milano, Via Celoria 16, Milano, I-20133, Italy; LASA, Department of Physics, Università Degli Studi di Milano and INFN-Milano, Via F.lli Cervi 201, Segrate, MI, I-20090, Italy. Electronic address:
Background: Knowledge on Bi metabolism in laboratory animals refers to studies at "extreme" exposures, i.e. pharmacologically relevant high-doses (mg kg b.
View Article and Find Full Text PDFACS Omega
January 2020
Departament de Bioquímica i Biologia Molecular, Estructura de Recerca Interdisciplinar en Biotecnologia i Biomedicina (ERI BioTecMed), Universitat de València. E-46100 Burjassot, Spain.
A peptide corresponding to bacteriorhodopsin (bR) helix C, later named pHLIP, inserts across lipid bilayers as a monomeric α-helix at acidic pH, but is an unstructured surface-bound monomer at neutral pH. As a result of such pH-responsiveness, pHLIP targets acidic tumors and has been used as a vehicle for imaging and drug-delivery cargoes. To gain insights about the insertion of bR helix C into biological membranes, we replaced two key aspartic residues that control the topological transition from the aqueous phase into a lipid bilayer.
View Article and Find Full Text PDFJ Biol Chem
March 2019
From the Department of Biochemistry and Biophysics, Stockholm University 10691 Stockholm, Sweden and
Astrotactin 1 (Astn1) and Astn2 are membrane proteins that function in glial-guided migration, receptor trafficking, and synaptic plasticity in the brain as well as in planar polarity pathways in the skin. Here we used glycosylation mapping and protease protection approaches to map the topologies of mouse Astn1 and Astn2 in rough microsomal membranes and found that Astn2 has a cleaved N-terminal signal peptide, an N-terminal domain located in the lumen of the rough microsomal membranes (topologically equivalent to the extracellular surface in cells), two transmembrane helices, and a large C-terminal lumenal domain. We also found that Astn1 has the same topology as Astn2, but we did not observe any evidence of signal peptide cleavage in Astn1.
View Article and Find Full Text PDFFEMS Yeast Res
November 2018
School of Life Sciences, Jawaharlal Nehru University, New Delhi-110 067, India.
Candida albicans N-acetylglucosaminylphosphatidylinositol de-N-acetylase (CaGpi12) recognises N-acetylglucosaminylphosphatidylinositol (GlcNAc-PI) from Saccharomyces cerevisiae and is able to complement ScGPI12 function. Both N- and C-terminal ends of CaGpi12 are important for its function. CaGpi12 was biochemically characterised using rough endoplasmic reticulum microsomes prepared from BWP17 strain of C.
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