Lysine peroxycarbamates: free radical-promoted peptide cleavage

Strategies are reported that combine in one step a predictable chemical-based protein digestion with mass spectrometry. Lysine residue amino groups in peptides and proteins are modified by reaction with a peroxycarbonate derived from p-nitrophenol, and tert-butyl hydroperoxide. The peroxycarbonate reacts with lysine residues in peptides and proteins, and the resulting lysine peroxycarbamates undergo homolytic fragmentation under conditions of low-energy collision-induced dissociation (CID). Observed fragmentation of the peptides involves apparent free radical processes including Hofmann-L?ffler-type rearrangements that lead to peptide chain fragmentation. Strategies for directed cleavage of peptides by free radical promoted processes are feasible, and such strategies may well simplify schemes for protein analysis.     

Diastereoselective free radical halogenation,azidation, and rearrangement of beta-silyl Barton esters

[reaction: see text] Barton esters of beta-silylcarboxylic acids were decomposed by photolysis alone in organic solvents or in the presence of ethanesulfonyl azide or bromotrichloromethane. Products of the reaction, beta-silylthiopyridyl ethers, beta-silyl azides, or alkenes, were formed with significant control of stereochemistry.     

Elastic scattering of positive pions from 12C at 30, 40 and 50 MeV

We have measured the angular distribution of π⁺ elastically scattered from ${}^{12}\text{C at}\text{E}\text{?}{}_{\mathrm{\pi }}\text{= 28.4, 38.6}\text{and}\text{48.9}\text{MeV}$. The angular range was between 13° and 165° lab. Scattered pions were detected with two counter telescopes which required pion identification through the observation of the decay muon. Data agree well with calculations derived from the free πn scattering amplitudes when absorption effects have been included.     

Multivariate statistical identification of human bladder carcinomas using ambient ionization imaging mass spectrometry

Diagnosis of human bladder cancer in untreated tissue sections is achieved by using imaging data from desorption electrospray ionization mass spectrometry (DESI-MS) combined with multivariate statistical analysis. We use the distinctive DESI-MS glycerophospholipid (GP) mass spectral profiles to visually characterize and formally classify twenty pairs (40 tissue samples) of human cancerous and adjacent normal bladder tissue samples. The individual ion images derived from the acquired profiles correlate with standard histological hematoxylin and eosin (H&E)-stained serial sections. The profiles allow us to classify the disease status of the tissue samples with high accuracy as judged by reference histological data. To achieve this, the data from the twenty pairs were divided into a training set and a validation set. Spectra from the tumor and normal regions of each of the tissue sections in the training set were used for orthogonal projection to latent structures (O-PLS) treated partial least-square discriminate analysis (PLS-DA). This predictive model was then validated by using the validation set and showed a 5% error rate for classification and a misclassification rate of 12%. It was also used to create synthetic images of the tissue sections showing pixel-by-pixel disease classification of the tissue and these data agreed well with the independent classification that uses histological data by a certified pathologist. This represents the first application of multivariate statistical methods for classification by ambient ionization although these methods have been applied previously to other MS imaging methods. The results are encouraging in terms of the development of a method that could be utilized in a clinical setting through visualization and diagnosis of intact tissue.     

Effect of Pr-Ca substitution on the transport and magnetic behavior of LaMnO3 perovskite

The effect of simultaneous substitution of a fluctuating cation and a divalent cation in LaMnO₃ perovskite modifies the properties of the material to exhibit large valence colossal magnetoresistance (CMR) effect. A good example of these properties is (La_1−2x Pr _x Ca _x )MnO₃ (LPCMO) type CMR material. In this communication it is reported that, with the increase in x (for x=0.1, 0.15, 0.2), the T _c varies between 100 and 120 K with improvisation in metal-insulator transition. Interestingly, resistance increases with x from few hundred ohms to few kilo ohms with corresponding decrease in the unit cell volume. The results of the studies using X-ray diffraction (XRD), electrical resistivity, magnetoresistance and ac susceptibility measurements on LPCMO samples for understanding the structural, transport and magnetic properties are discussed in detail.     

Free radical-induced site-specific peptide cleavage in the gas phase: Low-energy collision-induced dissociation in ESI- and MALDI mass spectrometry

Protein identification is routinely accomplished by peptide sequencing using mass spectrometry (MS) after enzymatic digestion. Site-specific chemical modification may improve peptide ionization efficiency or sequence coverage in mass spectrometry. We report herein that amino group of lysine residue in peptides can be selectively modified by reaction with a peroxycarbonate and the resulting lysine peroxycarbamates undergo homolytic fragmentation under conditions of low-energy collision-induced dissociation (CID) in electrospray ionization (ESI) and matrix-assisted laser desorption and ionization (MALDI) MS. Selective modification of lysine residue in peptides by our strategy can induce specific peptide cleavage at or near the lysine site. Studies using deuterated analogues of modified lysine indicate that fragmentation of the modified peptides involves apparent free-radical processes that lead to peptide chain fragmentation and side-chain loss. The formation of a-, c-, or z-types of ions in MS is reminiscent of the proposed free-radical mechanisms in low-energy electron capture dissociation (ECD) processes that may have better sequence coverage than that of the conventional CID method. This site-specific cleavage of peptides by free radical- promoted processes is feasible and such strategies may aid the protein sequencing analysis and have potential applications in top-down proteomics.     

The 14C(α, α')14C and 13C(d,p)14C reactions

The isoscalar transition rates and neutron-stripping probabilities to states of ¹⁴C have been measured using the 35 MeV ¹⁴C(α, α')¹⁴C and 17.7 MeV ¹³C(d, p)¹⁴C reactions. States showing great charge asymmetries in pion scattering at 8.32 MeV (2⁺) and 11.7 MeV (4^?) were examined in detail. Isoscalar transition rates B(02) were determined to be 168, 96 and 74 fm⁴ for the 7.01, 8.32 and 10.45 MeV 2⁺ states, with identical single-neutron spectroscopic factors of 0.065, from the (d, p) data, for the lowest two states.     

Proton radii determinations from the ratio of π+ elastic scattering from 11B and 12C

A new method using low energy π⁺ elastic scattering is employed to determine the proton matter distribution of a nucleus relative to an adjacent nucleus differing by one proton. The difference in the rms charge radii of ¹²C and ¹¹B was determined to be 0.072 ± 0.021 fm with π⁺ beam energies of 38.6 and 47.7 MeV.     

Elastic scattering of π− from 12C at 29 MeV

We have measured the differential cross section for π^? elastically scattered from ${}^{12}\text{C at}\text{T}\text{?}{}_{\mathrm{\pi }}\text{=29}\text{MeV}$. The scattered π^? were detected with pion range telescopes. The measured angular distribution is compared with a momentum space optical potential calculation. Derived phase shifts are compared with those obtained from π⁺ data and from momentum and coordinate space calculations.