Identification of Host Proteins Predictive of Early Stage Mycobacterium tuberculosis Infection.

The objective of this study was to identify blood-based protein biomarkers of early stage Mycobacterium tuberculosis (Mtb) infection. We utilized plasma and serum specimens from TB patients and their contacts (age≥12) enrolled in a household contact study in Uganda. In the discovery phase cross-sectional samples from 104 HIV-uninfected persons classified as either active TB, latent Mtb infection (LTBI), tuberculin skin test (TST) converters, or persistent TST-negative were analyzed. Two hundred eighty-nine statistically significant (false discovery rate corrected p<0.05) differentially expressed proteins were identified across all comparisons. Proteins associated with cellular immunity and lipid metabolism were induced early after Mtb infection. One hundred and fifty-nine proteins were selected for a targeted mass spectrometry assay. A set of longitudinal samples from 52 TST-negative subjects who converted to TST-positive or remained TST-negative were analyzed, and multivariate logistic regression was used to identify unique protein panels able to predict TST conversion with cross-validated AUC>0.85. Panel performance was confirmed with an independent validation set of longitudinal samples from 16 subjects. These candidate protein biomarkers may allow for the identification of recently Mtb infected individuals at highest risk for developing active TB and most likely to benefit from preventive therapy.

Fragmentation chemistry of [Met-Gly]•+, [Gly-Met]•+, and [Met-Met]•+ radical cations.

Radical cations [Met-Gly](•+), [Gly-Met](•+), and [Met-Met](•+) have been generated through collision-induced dissociation (CID) of [Cu(II)(CH3CN)2(peptide)](•2+) complexes. Their fragmentation patterns and dissociation mechanisms have been studied both experimentally and theoretically using density functional theory at the UB3LYP/6-311++G(d,p) level. The captodative structure, in which the radical is located at the α-carbon of the N-terminal residue and the proton is on the amide oxygen, is the lowest energy structure on each potential energy surface. The canonical structure, with the charge and spin both located on the sulfur, and the distonic ion with the proton on the terminal amino group, and the radical on the α-carbon of the C-terminal residue have similar energies. Interconversion between the canonical structures and the captodative isomers is facile and occurs prior to fragmentation. However, isomerization to produce the distonic structure is energetically less favorable and cannot compete with dissociation except in the case of [Gly-Met](•+). Charge-driven dissociations result in formation of [b(n) - H](•+) and a(1) ions. Radical-driven dissociation leads to the loss of the side chain of methionine as CH3-S-CH=CH2 producing α-glycyl radicals from both [Gly-Met](•+) and [Met-Met](•+). For [Met-Met](•+), loss of the side chain occurs at the C-terminal as shown by both labeling experiments and computations. The product, the distonic ion of [Met-Gly](•+), NH3 (+)CH(CH2CH2SCH3)CONHCH(•)COOH dissociates by loss of CH3S(•). The isomeric distonic ion NH3 (+)CH2CONHC(•)(CH2CH2SCH3)COOH is accessible directly from the canonical [Gly-Met](•+) ion. A fragmentation pathway that characterizes this ion (and the distonic ion of [Met-Met](•+)) is homolytic fission of the Cß-Cγ bond to lose CH3SCH2(•).