Recent Developments in Structural Proteomics: From Protein Identifications and Structure Determinations to Protein-Protein Interactions

Juan, Hsueh‐Fen; Liu, Hsuan-Liang; Hsu, Jyh‐Ping

doi:10.2174/1570164043379208

Cited by 2 publications

(2 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ith tremendous advances in proteomics in the past decade, the interest in understanding protein-protein interactions at a molecular level has expanded as new methods for exploring macromolecules have emerged [1][2][3]. Chemical crosslinking of proteins coupled with mass spectrometric analysis has become a more widely used method for determining protein-protein interactions in recent years [4][5][6].…”

mentioning

confidence: 99%

Impact of proline and aspartic acid residues on the dissociation of intermolecularly crosslinked peptides

Gardner

Brodbelt

2008

J. Am. Soc. Mass Spectrom.

View full text Add to dashboard Cite

The dissociation of intermolecularly crosslinked peptides was evaluated for a series of peptides with proline or aspartic acid residues positioned adjacent to the crosslinking sites (lysine residues). The peptides were crosslinked with either disuccinimidyl suberate (DSS) or disuccinimidyl L-tartrate (DST), and the influence of proline and aspartic acid residues on the fragmentation patterns were investigated for precursor ions with and without a mobile proton. Collisionally activated dissociation (CAD) spectra of aspartic acid-containing crosslinked peptide ions, doublycharged with both protons sequestered, were dominated by cleavage C-terminal to the Asp residue, similar to that of unmodified peptides. The proline-containing crosslinked peptides exhibited a high degree of internal ion formation, with the resulting product ions having an N-terminal proline residue. Upon dissociation of the doubly-charged crosslinked peptides, twenty to fifty percent of the fragment ion abundance was accounted for by multiple cleavage products. Crosslinked peptides possessing a mobile proton yielded almost a full series of b-and y-type fragment ions, with only proline-directed fragments still observed at high abundances. Interestingly, the crosslinked peptides exhibited a tendency to dissociate at the amide bond C-terminal to the crosslinked lysine residue, relative to the N-terminal side. One could envision updating computer algorithms to include these crosslinker specific product ions-particularly for precursor ions with localized protons-that provide complementary and confirmatory information, to offer more confident identification of both the crosslinked peptides and the location of the crosslink, as well as affording predictive guidelines for interpretation of the product-ion spectra of crosslinked peptides. (J

show abstract

mentioning

confidence: 99%

Impact of proline and aspartic acid residues on the dissociation of intermolecularly crosslinked peptides

Gardner

Brodbelt

2008

J. Am. Soc. Mass Spectrom.

View full text Add to dashboard Cite

show abstract

“…Since at this point neither the mode of pioglitazone binding to mitoNEET nor the mechanism of the stabilization of the Fe-S cluster of mitoNEET by pioglitazone binding have been elucidated, continued structure-function studies are needed, particularly using the well established methods of NMR spectroscopy and X-ray crystallography (Juan et al, 2004;Singh et al, 2006; for a review, see Burley & Bonanno, 2002). For future studies, we desire a robust method for the expression and isolation of mutant or full-length mitoNEET.…”

Section: Introductionmentioning

confidence: 99%

The novel 2Fe–2S outer mitochondrial protein mitoNEET displays conformational flexibility in its N-terminal cytoplasmic tethering domain

Conlan

Paddock

Axelrod

et al. 2009

Acta Crystallogr F Struct Biol Cryst Commun

View full text Add to dashboard Cite

A primary role for mitochondrial dysfunction is indicated in the pathogenesis of insulin resistance. A widely used drug for the treatment of type 2 diabetes is pioglitazone, a member of the thiazolidinedione class of molecules. MitoNEET, a 2Fe-2S outer mitochondrial membrane protein, binds pioglitazone [Colca et al. (2004), Am. J. Physiol. Endocrinol. Metab. 286, E252-E260]. The soluble domain of the human mitoNEET protein has been expressed C-terminal to the superfolder green fluorescent protein and the mitoNEET protein has been isolated. Comparison of the crystal structure of mitoNEET isolated from cleavage of the fusion protein (1.4 Å resolution, R factor = 20.2%) with other solved structures shows that the CDGSH domains are superimposable, indicating proper assembly of mitoNEET. Furthermore, there is considerable flexibility in the position of the cytoplasmic tethering arms, resulting in two different conformations in the crystal structure. This flexibility affords multiple orientations on the outer mitochondrial membrane.

show abstract

Recent Developments in Structural Proteomics: From Protein Identifications and Structure Determinations to Protein-Protein Interactions

Cited by 2 publications

References 0 publications

Impact of proline and aspartic acid residues on the dissociation of intermolecularly crosslinked peptides

Impact of proline and aspartic acid residues on the dissociation of intermolecularly crosslinked peptides

The novel 2Fe–2S outer mitochondrial protein mitoNEET displays conformational flexibility in its N-terminal cytoplasmic tethering domain

Contact Info

Product

Resources

About