Durand, Kirt Lenroy. Ph.D., Purdue University, December 2014. Determination of Disulfide Bond Connecting Patterns via Tandem Mass Spectrometry (MS) and Biomolecular Ion/Radical Reactions. Major Professor: Yu Xia. Disulfide bond formation is one of the most common post translational modifications to occur in proteins and naturally occurring peptides. Disulfide bond formation plays a critical rol...

The copper chaperone for superoxide dismutase 1 (Ccs1) provides an important cellular function against oxidative stress. Ccs1 is present in the cytosol and in the intermembrane space (IMS) of mitochondria. Its import into the IMS depends on the Mia40/Erv1 disulfide relay system, although Ccs1 is, in contrast to typical substrates, a multidomain protein and lacks twin Cx(n)C motifs. We report on...

Disulfide oxidoreductases are ubiquitous proteins in eukaryotes and prokaryotes. They catalyze the in vivo formation of disulfide bonds, which is critical for the stability and activity of many proteins. In E. coli, disulfide bond formation occurs in the periplasm and is primarily catalyzed through the actions of the highly oxidizing soluble proteinDsbA [1]. Homologues ofDsbA are found in many ...

Disulfide bonds are required for the stability and function of a large number of proteins. Recently, the results from genome analysis have suggested an important role for disulfide bonds concerning the structural stabilization of intracellular proteins from hyperthermophilic Archaea and Bacteria, contrary to the conventional view that structural disulfide bonds are rare in proteins from Archaea...

To study the importance of individual sulfhydryl residues during the folding and assembly in vivo of influenza virus hemagglutinin (HA), we have constructed and expressed a series of mutant HA proteins in which cysteines involved in three disulfide bonds have been substituted by serine residues. Investigations of the structure and intracellular transport of the mutant proteins indicate that (a)...

Human fibrinogen is a dimer, and each half-molecule is composed of three different polypeptides (A alpha, B beta, and gamma). The two half-molecules are joined together at the amino-terminal ends by three symmetrical disulfide bonds between adjacent A alpha chains, at position 28, and between adjacent gamma chains at gamma 8 and 9. To determine the role of these disulfide bonds in fibrinogen as...

Disulfide bonds play an important role in understanding protein folding, evolution, and in studies related to determining structural and functional properties of specific proteins. At the state-of-the-art, a large number of computational techniques have been proposed for determining disulfide bonds. Operating across the gamut of input data, from pure sequence-based information to spectra from m...

Disulfide bonds generally show only limited cleavage in positive ion mode collision activated dissociation (CAD). However, it has been emonstrated that a reverse situation exists in negative ion mode in which preferential S S and C S bond cleavage occurs. Here, we show that lectron detachment dissociation (EDD) and infrared multiphoton dissociation (IRMPD) of peptide anions containing disulfide...

Introduction of disulfide bonds into proteins entering the secretory pathway is catalyzed by Ero1p, which generates disulfide bonds de novo, and Pdi1p, which transfers disulfides to substrate proteins. A sufficiently oxidizing environment must be maintained in the endoplasmic reticulum (ER) to allow for disulfide formation, but a pool of reduced thiols is needed for isomerization of incorrectly...

The problem of protein structure prediction (PSP) is one of the main challenges in structural bioinformatics. To tackle this problem, PSP can be divided into several subproblems. One of these subproblems is the prediction of disulfide bonds. The disulfide connectivity prediction problem consists in identifying which nonadjacent cysteines would be cross-linked from all possible candidates. Deter...