Abstract:
:Autophagy is an evolutionarily conserved lysosome-mediated degradation and recycling process, which functions in cellular homeostasis and stress adaptation. The process is highly dynamic and involves autophagosome synthesis, cargo recognition and transport, autophagosome-lysosome fusion, and cargo degradation. The multistep nature of autophagy makes it challenging to quantify, and it is important to consider not only the number of autophagosomes within a cell but also the autophagic degradative activity. The rate at which cargos are recognized, segregated, and degraded through the autophagy pathway is defined as autophagic flux. In practice, methods to measure autophagic flux typically evaluate the lysosome-mediated cargo degradation step by leveraging known autophagy markers such as MAP1LC3B (microtubule-associated proteins 1A/1B light chain 3 beta) or lysosome-dependent fluorescent agents. In this review, we summarize the tools and methods used in mammalian cultured cells pertaining to these two approaches, and highlight innovations that have led to their evolution in recent years. We also discuss the potential limitations of these approaches and recommend using a combination of strategies and multiple different autophagy markers to reliably evaluate autophagic flux in mammalian cells.
journal_name
Biochem Soc Transjournal_title
Biochemical Society transactionsauthors
Yang KC,Sathiyaseelan P,Ho C,Gorski SMdoi
10.1042/BST20170102subject
Has Abstractpub_date
2018-02-19 00:00:00pages
97-110issue
1eissn
0300-5127issn
1470-8752pii
BST20170102journal_volume
46pub_type
杂志文章,评审abstract::The central role of VEGF (vascular endothelial growth factor A) in angiogenesis is dependent upon its ability to co-ordinately regulate multiple endothelial functions. The multifunctionality of VEGF at the cellular level results from its ability to initiate a diverse, complex and integrated network of signalling pathw...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/bst0311171
更新日期:2003-12-01 00:00:00
abstract::Senescence is a tumour suppressor mechanism which is cell-intrinsically activated in the context of cellular stress. Senescence can further be propagated to neighbouring cells, a process called secondary senescence induction. Secondary senescence was initially shown as a paracrine response to the secretion of cytokine...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST20190109
更新日期:2020-06-30 00:00:00
abstract::Isolation and purification of the [NiFe] hydrogenase of Desulfovibrio vulgaris Miyazaki F under aerobic conditions leads to a mixture of two states, Ni-A (unready) and Ni-B (ready). The two states are distinguished by different activation times and different EPR spectra. HYSCORE and ENDOR data and DFT calculations sho...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST0330007
更新日期:2005-02-01 00:00:00
abstract::Virtually all the biological processes that occur inside or outside cells are mediated by protein-protein interactions (PPIs). Hence, the charting and description of the PPI network, initially in organisms, the interactome, but more recently in specific tissues, is essential to fully understand cellular processes both...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20150001
更新日期:2016-06-15 00:00:00
abstract::The neurotrophins nerve growth factor, brain-derived neurotrophic factor, NT-3 (neurotrophin 3) and NT-4 are known for regulating neuron development, function and survival. Beyond this, neurotrophins were found to exert multiple effects on non-neuronal cells such as immune cells, smooth muscle and epithelial cells. In...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST0340594
更新日期:2006-08-01 00:00:00
abstract::Among the several signalling pathways regulated by ubiquitin and ubiquitin-like proteins, the one activating NF-kappaB (nuclear factor kappaB) is certainly one of the best characterized. The regulation of the activity of this transcription factor by members of the ubiquitin family occurs at various levels, imposing ov...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST0360853
更新日期:2008-10-01 00:00:00
abstract::Phosphorus is one of the most important macronutrients and is indispensable for all organisms as a critical structural component as well as participating in intracellular signalling and energy metabolism. Sensing and signalling of phosphate (Pi) has been extensively studied and is well understood in single-cellular or...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20160026
更新日期:2016-06-15 00:00:00
abstract::Intracellular pathogens survive in eukaryotic cells by evading a variety of host defences. To avoid degradation through the endocytic pathway, intracellular bacteria must adapt their phagosomes into protective compartments that promote bacterial replication. Legionella pneumophila and Coxiella burnetii are Gram-negati...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20120167
更新日期:2012-12-01 00:00:00
abstract::Phosphoinositide 3-kinases (PI3Ks) are important signalling enzymes in most cell types. Recent gene targeting studies have shed light on the importance of this family of lipid kinases in the immune system, and the complex mechanisms by which these kinases are regulated in vivo. We have recently reported a phenotype of...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/bst0310270
更新日期:2003-02-01 00:00:00
abstract::Three mammalian expression systems suitable for expressing recombinant receptors have been described. Each is suited to a different aspect of the study of receptors and their behaviour. IRES-based vectors are ideal for creating stable mammalian cell lines suitable for screening receptors using a signalling readout. Un...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/bst0260699
更新日期:1998-11-01 00:00:00
abstract::Reciprocal biochemical and biophysical interactions between tumor cells, stromal cells and the extracellular matrix (ECM) result in a unique tumor microenvironment that determines disease outcome. The cellular component of the tumor microenvironment contributes to tumor growth by providing nutrients, assisting in the ...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20180334
更新日期:2019-02-28 00:00:00
abstract::Here, we describe a recently discovered O-GlcNAc transferase termed EOGT for EGF domain-specific O-GlcNAc transferase. EOGT transfers GlcNAc (N-acetylglucosamine) to Ser or Thr in secreted and membrane proteins that contain one or more epidermal growth factor-like repeats with a specific consensus sequence. Thus, EOGT...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20160165
更新日期:2017-04-15 00:00:00
abstract::Many plant viral RNAs lack the 5'-cap structure that is required on all host mRNAs for interacting with essential translation initiation factors. Instead, uncapped viral RNAs take over the host translation machinery by harbouring sequences that functionally replace the 5'-cap. Recent reports reveal at least eight diff...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST0351629
更新日期:2007-12-01 00:00:00
abstract::In this study the network of ATP-consuming and -producing reactions, interacting via the cytosolic ATP/ADP+P(i) system, was studied for the first time in an intact organ, the isolated perfused rat liver, using top-down metabolic-control analysis. Flux control in the metabolically resting state (only oxidative phosphor...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/bst0280109
更新日期:2000-02-01 00:00:00
abstract::Non-alcoholic fatty liver disease (NAFLD) is one of the most common liver disorders in industrialized countries. NAFLD develops in the absence of alcohol abuse and encompasses a wide spectrum of disorders ranging from benign fatty liver to non-alcoholic steatohepatitis (NASH). NASH often leads to fibrosis, cirrhosis a...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20140138
更新日期:2014-10-01 00:00:00
abstract::The GABAR [GABA(A) (gamma-aminobutyric acid type A) receptor], which mediates most inhibition in the brain, is regulated homoeostatically to maintain an optimal level of neuronal excitability. In particular, the alpha(4)betadelta subtype of the GABAR plays a pivotal role in this regulation. This receptor, which is exp...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST0371378
更新日期:2009-12-01 00:00:00
abstract::HD (Huntington's disease) is caused by a polyQ (polyglutamine) expansion in the huntingtin protein, which leads to protein misfolding and aggregation of this protein. Abnormal copper accumulation in the HD brain was first reported more than 15 years ago. Recent findings show that copper-regulatory genes are induced du...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST0380552
更新日期:2010-04-01 00:00:00
abstract::RAMPs (receptor activity-modifying proteins) are single-pass transmembrane proteins that associate with certain family-B GPCRs (G-protein-coupled receptors). Specifically for the CT (calcitonin) receptor-like receptor and the CT receptor, this results in profound changes in ligand binding and receptor pharmacology, al...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST0320843
更新日期:2004-11-01 00:00:00
abstract::An RNA world has been placed centre stage for explaining the origin of life. Indeed, RNA is the most plausible molecule able to form both a (self)-replicator and to inherit information, necessities for initiating genetics. However, in parallel with self-replication, the proto-organism had to obtain the ability to cata...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20140132
更新日期:2014-08-01 00:00:00
abstract::Structure-based design has gained credibility as a valuable component of the modern drug discovery process. The technique of SSNMR (solid-state NMR) promises to be a useful counterpart to the conventional experimental techniques of X-ray crystallography and solution-state NMR for providing structural features of drug ...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST0350985
更新日期:2007-11-01 00:00:00
abstract::We have identified two basic residues that are important for the recognition of secretin and vasoactive intestinal peptide (VIP) by their respective receptors. These two peptides containing an Asp residue at position 3 interacted with an arginine residue in transmembrane helix 2 (TM2) of the receptor, and the lysine r...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/bst0300437
更新日期:2002-08-01 00:00:00
abstract::The inability of the human heart to effectively repair itself after acute ischaemic injury has driven the search for efficacious means of promoting cardiac regenerative growth. Central to this has been the emergence of cell-based strategies to stimulate and augment both myocardial regeneration and neovascularization. ...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST0371218
更新日期:2009-12-01 00:00:00
abstract::We have been analysing the signalling systems that couple to receptors of the TIR (Toll/interleukin-1 receptor) family, which signal through a common cytoplasm region; the TIR domain. These systems are of both practical and fundamental biological significance, being central to the pathogenesis of chronic inflammatory ...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/bst0311462
更新日期:2003-12-01 00:00:00
abstract::Increased production of amyloid β-peptide (Aβ) and altered processing of tau in Alzheimer's disease (AD) are associated with synaptic dysfunction, neuronal death and cognitive and behavioural deficits. Neuroinflammation is also a prominent feature of AD brain and considerable evidence indicates that inflammatory event...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST20140155
更新日期:2014-10-01 00:00:00
abstract::AMPK (AMP-activated protein kinase) is a cellular energy sensor that monitors the ratio of AMP/ATP, and possibly also ADP/ATP, inside cells. Once activated by falling cellular energy levels, it acts to restore energy homoeostasis by switching on catabolic pathways that generate ATP, while switching off anabolic pathwa...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST20130244
更新日期:2014-02-01 00:00:00
abstract::Bone morphogenetic proteins (BMPs) belong to the transforming growth factor-β (TGF-β) family signalling pathway. Similar to TGF-β, the complex roles of BMPs in development and disease are demonstrated by their dichotomous roles in various cancers and cancer stages. Although early studies implicated BMP signalling in t...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20160069
更新日期:2016-08-15 00:00:00
abstract::Mouse ES (embryonic stem) cells maintain pluripotency with robust proliferation in vitro. ES cells share some similarities with cancer cells, such as anchorage-independent growth, loss of contact inhibition and tumour formation. After differentiation, ES cells lose pluripotency and tumorigenicity. Recent studies showe...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20051522
更新日期:2005-12-01 00:00:00
abstract::PGs (prostaglandins) are synthesized through the cyclo-oxygenase (COX-1 and -2) pathway in a variety of cells in response to various physiological stimuli. All cells require at least one pathway for apoptosis, and mitochondrial play a central role in regulation of apoptosis. In a previous study, incubation of A549 cel...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST0321078
更新日期:2004-12-01 00:00:00
abstract::Photosynthesis is fundamental to biomass production, but is a dynamic process sensitive to environmental constraints. In recent years, approaches to increase biomass and grain yield by altering photosynthetically related processes in the plant have received considerable attention. However, improving biomass yield requ...
journal_title:Biochemical Society transactions
pub_type: 杂志文章,评审
doi:10.1042/BST20170296
更新日期:2018-04-17 00:00:00
abstract::Chromosomal instability occurs early in the development of cancer and may represent an important step in promoting the multiple genetic changes required for the initiation and/or progression of the disease. Telomere erosion is one of the factors that contribute to chromosome instability through end-to-end chromosome f...
journal_title:Biochemical Society transactions
pub_type: 杂志文章
doi:10.1042/BST0381698
更新日期:2010-12-01 00:00:00