NDLI: Alanine scan of core positions in ubiquitin reveals links between dynamics, stability, and function

Content Provider	PubMed Central
Author	Lee, Shirley Y. Pullen, Lester Virgil, Daniel J. Castañeda, Carlos A. Abeykoon, Dulith Bolon, Daniel N. A. David, Fushman
Copyright Year	2013
Abstract	Mutations at solvent inaccessible core positions in proteins can impact function through many biophysical mechanisms including alterations to thermodynamic stability and protein dynamics. As these properties of proteins are difficult to investigate, the impacts of core mutations on protein function are poorly understood for most systems. Here, we determined the effects of alanine mutations at all 15 core positions in ubiquitin on function in yeast. The majority (13 of 15) of alanine substitutions supported yeast growth as the sole ubiquitin. The two null mutants (I30A and L43A) were both less stable to temperature-induced unfolding in vitro than wild-type, but were well folded at physiological temperatures. Heteronuclear NMR studies indicated that the L43A mutation reduces temperature stability while retaining a ground-state structure similar to wild-type. This structure enables L43A to bind to common ubiquitin receptors in vitro. Many of the core alanine ubiquitin mutants, including one of the null variants (I30A), exhibited an increased accumulation of high molecular weight species, suggesting that these mutants caused a defect in the processing of ubiquitin-substrate conjugates. In contrast, L43A exhibited a unique accumulation pattern with reduced levels of high molecular weight species and undetectable levels of free ubiquitin. When conjugation to other proteins was blocked, L43A ubiquitin accumulated as free ubiquitin in yeast. Based on these findings we speculate that ubiquitin's stability to unfolding may be required for efficient recycling during proteasome-mediated substrate degradation.
Related Links	http://dx.doi.org/10.1016/j.jmb.2013.10.042
Ending Page	1389
Page Count	13
Starting Page	1377
File Format	PDF
ISSN	00222836
e-ISSN	10898638
Journal	Journal of molecular biology
Issue Number	7
Volume Number	426
Language	English
Publisher Date	2014-04-01
Access Restriction	Open
Subject Keyword	Molecular Biology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Structural Biology Molecular Biology Biophysics

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Molecular Links between the E2 Envelope Glycoprotein and Nucleocapsid Core in Sindbis Virus

NEDD8 links Cullin–Ring ubiquitin Ligase function to the p97 pathway

Alanine Scan of α-Conotoxin RegIIA Reveals a Selective α3β4 Nicotinic Acetylcholine Receptor Antagonist

Alanine Scanning Mutagenesis of Anti-TRAP (AT) Reveals Residues Involved in Binding to TRAP

Mutations in the Hydrophobic Core of Ubiquitin Differentially Affect its Recognition by Receptor Proteins

Conformational Dynamics and Structural Plasticity Play Critical Roles in Ubiquitin Recognition of a UIM Domain

Global Dynamics of Proteins: Bridging Between Structure and Function

Crosstalk between the Protein Surface and Hydrophobic Core in a Core-swappedFibronectin Type III Domain

Structural Bases for Stability-Function Tradeoffs in Antibiotic Resistance

Alanine scan of core positions in ubiquitin reveals links between dynamics, stability, and function

Similar Documents

Molecular Links between the E2 Envelope Glycoprotein and Nucleocapsid Core in Sindbis Virus

NEDD8 links Cullin–Ring ubiquitin Ligase function to the p97 pathway

Alanine Scan of α-Conotoxin RegIIA Reveals a Selective α3β4 Nicotinic Acetylcholine Receptor Antagonist

Alanine Scanning Mutagenesis of Anti-TRAP (AT) Reveals Residues Involved in Binding to TRAP

Mutations in the Hydrophobic Core of Ubiquitin Differentially Affect its Recognition by Receptor Proteins

Conformational Dynamics and Structural Plasticity Play Critical Roles in Ubiquitin Recognition of a UIM Domain

Global Dynamics of Proteins: Bridging Between Structure and Function

Crosstalk between the Protein Surface and Hydrophobic Core in a Core-swappedFibronectin Type III Domain

Structural Bases for Stability-Function Tradeoffs in Antibiotic Resistance

Alanine scan of core positions in ubiquitin reveals links between dynamics, stability, and function