Structure and Function of the Conserved Domain in αA-Crystallin. Site-Directed Spin Labeling Identifies a β-Strand Located near a Subunit Interface†

Biochemistry ◽  
1997 ◽  
Vol 36 (33) ◽  
pp. 9951-9957 ◽  
Author(s):  
Anderee R. Berengian ◽  
Michael P. Bova ◽  
Hassane S. Mchaourab
Keyword(s):  
Spin Labeling ◽  
Structure And Function ◽  
Conserved Domain ◽  
Subunit Interface ◽  
A Subunit ◽  
Site Directed Spin Labeling ◽  
And Function

scholarly journals Structure and Function of Synaptotagmin 1 C2 Domains as Determined by Site-Directed Spin Labeling

2011 ◽  
Vol 100 (3) ◽  
pp. 144a
Author(s):  
Dawn Z. Herrick ◽  
Weiwei Kuo ◽  
Jeffrey F. Ellena ◽  
David S. Cafiso
Keyword(s):  
Spin Labeling ◽  
Structure And Function ◽  
C2 Domains ◽  
Synaptotagmin 1 ◽  
Site Directed Spin Labeling ◽  
And Function

TBC proteins: GAPs for mammalian small GTPase Rab?

2011 ◽  
Vol 31 (3) ◽  
pp. 159-168 ◽  
Author(s):  
Mitsunori Fukuda
Keyword(s):  
Insulin Resistance ◽  
Small Gtpases ◽  
Structure And Function ◽  
Small Gtpase ◽  
Cell Cycle Regulators ◽  
Gtpase Activating Protein ◽  
Domain Family ◽  
Protein Motif ◽  
Conserved Domain ◽  
And Function

The TBC (Tre-2/Bub2/Cdc16) domain was originally identified as a conserved domain among the tre-2 oncogene product and the yeast cell cycle regulators Bub2 and Cdc16, and it is now widely recognized as a conserved protein motif that consists of approx. 200 amino acids in all eukaryotes. Since the TBC domain of yeast Gyps [GAP (GTPase-activating protein) for Ypt proteins] has been shown to function as a GAP domain for small GTPase Ypt/Rab, TBC domain-containing proteins (TBC proteins) in other species are also expected to function as a certain Rab-GAP. More than 40 different TBC proteins are present in humans and mice, and recent accumulating evidence has indicated that certain mammalian TBC proteins actually function as a specific Rab-GAP. Some mammalian TBC proteins {e.g. TBC1D1 [TBC (Tre-2/Bub2/Cdc16) domain family, member 1] and TBC1D4/AS160 (Akt substrate of 160 kDa)} play an important role in homoeostasis in mammals, and defects in them are directly associated with mouse and human diseases (e.g. leanness in mice and insulin resistance in humans). The present study reviews the structure and function of mammalian TBC proteins, especially in relation to Rab small GTPases.

Nanoscale lipid membrane mimetics in spin-labeling and electron paramagnetic resonance spectroscopy studies of protein structure and function

2017 ◽  
Vol 6 (1) ◽  
pp. 75-92 ◽  
Author(s):  
Elka R. Georgieva
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Protein Structure ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Spin Labeling ◽  
Structure And Function ◽  
Protein Structure And Function ◽  
Paramagnetic Resonance ◽  
And Function ◽  
Electron Paramagnetic

AbstractCellular membranes and associated proteins play critical physiological roles in organisms from all life kingdoms. In many cases, malfunction of biological membranes triggered by changes in the lipid bilayer properties or membrane protein functional abnormalities lead to severe diseases. To understand in detail the processes that govern the life of cells and to control diseases, one of the major tasks in biological sciences is to learn how the membrane proteins function. To do so, a variety of biochemical and biophysical approaches have been used in molecular studies of membrane protein structure and function on the nanoscale. This review focuses on electron paramagnetic resonance with site-directed nitroxide spin-labeling (SDSL EPR), which is a rapidly expanding and powerful technique reporting on the local protein/spin-label dynamics and on large functionally important structural rearrangements. On the other hand, adequate to nanoscale study membrane mimetics have been developed and used in conjunction with SDSL EPR. Primarily, these mimetics include various liposomes, bicelles, and nanodiscs. This review provides a basic description of the EPR methods, continuous-wave and pulse, applied to spin-labeled proteins, and highlights several representative applications of EPR to liposome-, bicelle-, or nanodisc-reconstituted membrane proteins.

Studies on the structure and function of the carp gonadotropin a subunit by site-directed mutagenesis

2009 ◽  
Vol 46 (3-4) ◽  
pp. 209-213 ◽  
Author(s):  
CHANG-JEN HUANG ◽  
FORE-LIEN HUANG ◽  
GEEN-DONG CHANG ◽  
YEA-SHA CHANG ◽  
TUNG-BIN LO
Keyword(s):  
Structure And Function ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
A Subunit ◽  
And Function

scholarly journals Accessing the distance range of interest in biomolecules: Site-directed spin labeling and DEER spectroscopy

2010 ◽  
Vol 24 (3-4) ◽  
pp. 283-288 ◽  
Author(s):  
Sabine Böhme ◽  
Heinz-Jürgen Steinhoff ◽  
Johann P. Klare
Keyword(s):  
Conformational Changes ◽  
Protein Interactions ◽  
Spin Labeling ◽  
Paramagnetic Resonance ◽  
Topological Information ◽  
Pulsed Epr ◽  
Site Directed Spin Labeling ◽  
Intermolecular Distances ◽  
And Function ◽  
Electron Paramagnetic

Investigations on the structure and function of biomolecules often depend on the availability of topological information to build up structural models or to characterize conformational changes during function. Electron paramagnetic resonance (EPR) spectroscopy in combination with site–directed spin labeling (SDSL) allow to determine intra- and intermolecular distances in the range from 4–70 Å, covering the range of interest for biomolecules. The approach does not require crystalline samples and is well suited also for molecules exhibiting intrinsic flexibility. This article is intended to give an overview on pulsed EPR in conjunction with SDSL to study protein interactions as well as conformational changes, exemplified on the tRNA modifying enzyme MnmE.

scholarly journals Posttranscriptional Site-Directed Spin Labeling of Large RNAs with an Unnatural Base Pair System Under Non-Denaturing Conditions

2020 ◽  
Author(s):  
Yan Wang ◽  
Venkatesan Kathiresan ◽  
Yaoyi Chen ◽  
Yanping Hu ◽  
Wei Jiang ◽  
...  
Keyword(s):  
Epr Spectroscopy ◽  
Base Pair ◽  
Continuous Wave ◽  
Double Resonance ◽  
Rnase P ◽  
Spin Labeling ◽  
Paramagnetic Resonance ◽  
Site Directed Spin Labeling ◽  
And Function

<div> <p>Site-directed spin labeling (SDSL) of large RNAs for electron paramagnetic resonance (EPR) spectroscopy remains challenging up-to-date. We here demonstrate an efficient and generally applicable posttranscriptional SDSL method for large RNAs under non-denaturing conditions using an expanded genetic alphabet containing the NaM-TPT3 unnatural base pair (UBP). An alkyne-modified TPT3 ribonucleotide triphosphate (rTPT3<sup>CO</sup>TP) is synthesized and site-specifically incorporated into large RNAs by <i>in vitro</i> transcription, which allows attachment of the azide-containing nitroxide through click chemistry. We validate this strategy using a 419-nucleotide Ribonuclease P (RNase P) RNA from Bacillus <i>stearothermophilus. </i>The effects of site-directed UBP incorporation and subsequent spin labeling to global structure and function of RNase P are marginal as evaluated by Circular Dichroism spectroscopy, Small Angle X-ray Scattering, and enzymatic assay. Continuous-wave EPR analyses reveal that the labeling reaction is efficient and specific, and Pulsed Electron-Electron Double Resonance measurements yield an inter-spin distance distribution that agrees well with the crystal structure. Thus, the labeling strategy as presented overcomes the size constraint of RNA labeling, opening new possibilities for application of EPR spectroscopy in investigating structure and dynamics of large RNA.</p> </div> <br>

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