We have recently developed and implemented two experiments in biomolecular NMR for an undergraduate-level biophysical chemistry laboratory with commercially available (15) N-enriched human ubiquitin. These experiments take advantage of (15) N direct detection of the NMR signal. The first experiment develops skills in acquiring and interpreting one-dimensional and two-dimensional NMR data of an aqueous protein sample (D. Rovnyak, L. E. Thompson (2005) An accessible two-dimensional solution nuclear magnetic resonance experiment on human ubiquitin, Biochem. Mol. Biol. Educ. 33, 117-122). In the second experiment, discussed here, students examine the overall dynamics of a protein in solution. Students measure spin relaxation times T(1) and T(2) and use these data to estimate a rotational correlation time, τ(c) , and the diameter of ubiquitin using the Stokes-Einstein relation. This is a fast and simple experiment, requiring about 1 h of instrument time, and a valuable opportunity to provide a hands-on experience in studying macromolecular size and dynamics. This experiment is conducted in synchrony with lecture material on hydrodynamics and sedimentation.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1002/bmb.23 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Protein quality control machinery: regulators of condensate architecture and functionality.
Trends Biochem Sci
January 2025
Department of Chemistry, Syracuse University, Syracuse, NY 13244, USA; Department of Biology, Syracuse University, Syracuse, NY 13244, USA; Bioinspired Institute, Syracuse University, Syracuse, NY 13244, USA; Interdisciplinary Neuroscience Program, Syracuse University, Syracuse, NY 13244, USA. Electronic address:
Protein quality control (PQC) mechanisms including the ubiquitin (Ub)-proteasome system (UPS), autophagy, and chaperone-mediated refolding are essential to maintain protein homeostasis in cells. Recent studies show that these PQC mechanisms are further modulated by biomolecular condensates that sequester PQC components and compartmentalize reactions. Accumulating evidence points towards the PQC machinery playing a pivotal role in regulating the assembly, disassembly, and viscoelastic properties of several condensates.
View Article and Find Full Text PDFParkin modulates the hepatocellular carcinoma microenvironment by regulating PD-1/PD-L1 signalling.
J Adv Res
January 2025
Cancer Center, Department of Medical Oncology, Key Laboratory of Tumor Molecular Diagnosis and Individualized Medicine of Zhejiang Province, Zhejiang Provincial People's Hospital (Affiliated People's Hospital, Hangzhou Medical College), Hangzhou, China. Electronic address:
Introduction: Parkin-mediated mitophagy is essential for the clearance of damaged mitochondria, and it inhibits tumour development. The role of mitophagy in modulating tumour immunity is becoming clearer, but the underlying mechanism is still poorly understood.
Objective: This study was designed to examine the role for Parkin in the immune microenvironment of liver tumors induced by carbon tetrachloride (CCl).
The efflux pump ABCC1/MRP1 constitutively restricts PROTAC sensitivity in cancer cells.
Cell Chem Biol
December 2024
CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, 1090 Vienna, Austria; Center for Physiology and Pharmacology, Medical University of Vienna, 1090 Vienna, Austria. Electronic address:
Proteolysis targeting chimeras (PROTACs) are bifunctional molecules that induce selective protein degradation by linking an E3 ubiquitin ligase enzyme to a target protein. This approach allows scope for targeting "undruggable" proteins, and several PROTACs have reached the stage of clinical candidates. However, the roles of cellular transmembrane transporters in PROTAC uptake and efflux remain underexplored.
View Article and Find Full Text PDFRole of TRIM29 in disease: What is and is not known.
Int Immunopharmacol
January 2025
Key Laboratory of Livestock Infectious Diseases, Ministry of Education, Key Laboratory of Zoonosis, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, 120 Dongling Road, Shenyang 110866, China; The Research Unit for Pathogenic Mechanisms of Zoonotic Parasites, Chinese Academy of Medical Sciences, 120 Dongling Road, Shenyang 110866, China. Electronic address:
Tripartite motif-containing proteins (TRIMs), comprising the greatest subfamily of E3 ubiquitin ligases with approximately 80 members of this family, are widely distributed in mammalian cells. TRIMs actively participate in ubiquitination of target proteins, a type of post-translational modification associated with protein degradation and other functions. Tripartite motif-containing protein 29 (TRIM29), a member of the TRIM family, differs from other members of this family in that it lacks the RING finger structural domain containing cysteine and histidine residues that mediates DNA binding, protein-protein interactions, and ubiquitin ligase, at its N-terminus.
View Article and Find Full Text PDFCalcium peroxide treatment of cyanobacterial blooms: Ecological safety assessment on submerged macrophyte Vallisneria natans.
Sci Total Environ
January 2025
Department of Environmental Science and Engineering, Fudan University, Shanghai, PR China.
The outbreak of cyanobacterial blooms poses an increasingly serious ecological challenge. Our previous study found that calcium peroxide (CaO) has a high inhibitory effect on cyanobacteria, along with a practical application potential in cyanobacteria-dominated lakes. In order to explore the sensitivity of aquatic ecosystems to CaO treatment, we conducted this study to elucidate the ecological impact of CaO on Vallisneria natans (V.
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!