Segmental Isotopic Labeling of Ubiquitin Chains To Unravel Monomer-Specific Molecular Behavior

2011 ◽  
Vol 123 (47) ◽  
pp. 11406-11410 ◽  
Author(s):  
Carlos A. Castañeda ◽  
Liat Spasser ◽  
Sudhir N. Bavikar ◽  
Ashraf Brik ◽  
David Fushman
Keyword(s):  
Isotopic Labeling ◽  
Molecular Behavior ◽  
Segmental Isotopic Labeling

scholarly journals Segmental Isotopic Labeling of Ubiquitin Chains To Unravel Monomer-Specific Molecular Behavior

2011 ◽  
Vol 50 (47) ◽  
pp. 11210-11214 ◽  
Author(s):  
Carlos A. Castañeda ◽  
Liat Spasser ◽  
Sudhir N. Bavikar ◽  
Ashraf Brik ◽  
David Fushman
Keyword(s):  
Isotopic Labeling ◽  
Molecular Behavior ◽  
Segmental Isotopic Labeling

Segmental Isotopic Labeling of a Central Domain in a Multidomain Protein by ProteinTrans-Splicing Using Only One Robust DnaE Intein

2009 ◽  
Vol 48 (33) ◽  
pp. 6128-6131 ◽  
Author(s):  
Alena E. L. Busche ◽  
A. Sesilja Aranko ◽  
Mehdi Talebzadeh-Farooji ◽  
Frank Bernhard ◽  
Volker Dötsch ◽  
...  
Keyword(s):  
Isotopic Labeling ◽  
Central Domain ◽  
Multidomain Protein ◽  
Segmental Isotopic Labeling

Segmental Isotopic Labeling of a Central Domain in a Multidomain Protein by ProteinTrans-Splicing Using Only One Robust DnaE Intein

2009 ◽  
Vol 121 (33) ◽  
pp. 6244-6247 ◽  
Author(s):  
Alena E. L. Busche ◽  
A. Sesilja Aranko ◽  
Mehdi Talebzadeh-Farooji ◽  
Frank Bernhard ◽  
Volker Dötsch ◽  
...  
Keyword(s):  
Isotopic Labeling ◽  
Central Domain ◽  
Multidomain Protein ◽  
Segmental Isotopic Labeling

scholarly journals Segmental isotopic labeling by asparaginyl endopeptidase-mediated protein ligation

2018 ◽  
Vol 71 (4) ◽  
pp. 225-235 ◽  
Author(s):  
Kornelia M. Mikula ◽  
Luisa Krumwiede ◽  
Andreas Plückthun ◽  
Hideo Iwaï
Keyword(s):  
Isotopic Labeling ◽  
Protein Ligation ◽  
Asparaginyl Endopeptidase ◽  
Segmental Isotopic Labeling

scholarly journals Effects of an HIV-1 maturation inhibitor on the structure and dynamics of CA-SP1 junction helices in virus-like particles

2020 ◽  
Vol 117 (19) ◽  
pp. 10286-10293 ◽  
Author(s):  
Sebanti Gupta ◽  
John M. Louis ◽  
Robert Tycko
Keyword(s):  
Chemical Shifts ◽  
Isotopic Labeling ◽  
Relaxation Rates ◽  
Virus Like Particles ◽  
Gag Polyprotein ◽  
Structure And Dynamics ◽  
Maturation Inhibitor ◽  
Rna Maturation ◽  
Segmental Isotopic Labeling ◽  
Hiv 1

HIV-1 maturation involves conversion of the immature Gag polyprotein lattice, which lines the inner surface of the viral membrane, to the mature capsid protein (CA) lattice, which encloses the viral RNA. Maturation inhibitors such as bevirimat (BVM) bind within six-helix bundles, formed by a segment that spans the junction between the CA and spacer peptide 1 (SP1) subunits of Gag, and interfere with cleavage between CA and SP1 catalyzed by the HIV-1 protease (PR). We report solid-state NMR (ssNMR) measurements on spherical virus-like particles (VLPs), facilitated by segmental isotopic labeling, that provide information about effects of BVM on the structure and dynamics of CA–SP1 junction helices in the immature lattice. Although BVM strongly blocks PR-catalyzed CA–SP1 cleavage in VLPs and blocks conversion of VLPs to tubular CA assemblies, 15N and 13C ssNMR chemical shifts of segmentally labeled VLPs with and without BVM are very similar, indicating that interaction with BVM does not alter the six-helix bundle structure appreciably. Only the 15N chemical shift of A280 (the first residue of SP1) changes significantly, consistent with BVM binding to an internal ring of hydrophobic side chains of L279 residues. Measurements of transverse 15N spin relaxation rates reveal a reduction in the amplitudes and/or timescales of backbone N–H bond motions, corresponding to a rigidification of the six-helix bundles. Overall, our data show that inhibition of HIV-1 maturation by BVM involves changes in structure and dynamics that are surprisingly subtle, but still sufficient to produce a large effect on CA–SP1 cleavage.

scholarly journals Combining DNP NMR with segmental and specific labeling to study a yeast prion protein strain that is not parallel in-register

2017 ◽  
Vol 114 (14) ◽  
pp. 3642-3647 ◽  
Author(s):  
Kendra K. Frederick ◽  
Vladimir K. Michaelis ◽  
Marc A. Caporini ◽  
Loren B. Andreas ◽  
Galia T. Debelouchina ◽  
...  
Keyword(s):  
Prion Protein ◽  
Long Range ◽  
Isotopic Labeling ◽  
Yeast Prion ◽  
Long Range Interactions ◽  
Intermolecular Contacts ◽  
Protein Strain ◽  
Segmental Isotopic Labeling ◽  
Chemical Shift Degeneracy

The yeast prion protein Sup35NM is a self-propagating amyloid. Despite intense study, there is no consensus on the organization of monomers within Sup35NM fibrils. Some studies point to a β-helical arrangement, whereas others suggest a parallel in-register organization. Intermolecular contacts are often determined by experiments that probe long-range heteronuclear contacts for fibrils templated from a 1:1 mixture of 13C- and 15N-labeled monomers. However, for Sup35NM, like many large proteins, chemical shift degeneracy limits the usefulness of this approach. Segmental and specific isotopic labeling reduce degeneracy, but experiments to measure long-range interactions are often too insensitive. To limit degeneracy and increase experimental sensitivity, we combined specific and segmental isotopic labeling schemes with dynamic nuclear polarization (DNP) NMR. Using this combination, we examined an amyloid form of Sup35NM that does not have a parallel in-register structure. The combination of a small number of specific labels with DNP NMR enables determination of architectural information about polymeric protein systems.

Sign in / Sign up

Export Citation Format

Share Document