Top-down proteomics (TDP) offers a powerful approach to identifying intact proteoforms, inherently providing information about protein modifications. Truncated proteoforms are among the most frequently observed modifications in TDP studies. Here, we selected fifty datasets, including over 140,000 proteoforms, published over the past decade, spanning various organisms, sample preparation...
Top-down proteomics could offer comprehensive investigations of proteoforms. However, it is greatly challenged by the co-elution of intact proteins which results in overlapped mass spectra. Hence, highly effective protein separation to reduce the co-elution of intact proteins from complex samples is of vital importance.
The ethane-bridged hybrid monolithic column with homogeneous macropores...
Phosphorylation-mediated signaling dynamics across spatially distinct cardiac regions remain poorly understood due to limited technical capacity for deep and sensitive analysis of minute samples. Here, we present an optimized TiO₂-based micropipette tip method for deep phosphoproteomics, achieving high sensitivity (12,117 class I phosphosites from only 10 µg HeLa peptides) and reproducibility....
Introduction
Top-down proteomics (TDP) is increasingly being applied in proteoform-resolved biomedical and clinical research. The complexity of TDP data demands flexible visualization tools integrated with analysis workflows to streamline interpretation and validation. Existing tools often lack adaptability and interactivity, requiring complimentary manual analysis to generate...
Cervical cancer remains a significant global health issue due to its high morbidity and mortality rates. Recently, Lactobacillus crispatus has been recognized for its crucial role in maintaining cervical health. While some studies have explored the use of L. crispatus to mitigate cervical cancer, the underlying mechanisms remain largely unknown. In this study, we employed non-targeted...
Hemoglobin is a tetrameric protein responsible for the blood oxygen transport. Any abnormalities in hemoglobin structure can lead to serious health outcomes. Although very well characterized in humans, it has been scarcely studied in the case of birds. Some studies have been carried out at the globin level without providing further evidences about the native structure of the complex. In this...
Targeted protein degradation of undruggable proteins is transformative in drug discovery. Molecular glues (MGs) enhance weak interactions between targets and E3 ligase. Native mass spectrometry (nMS) identifies E3-MG-target complexes directly, but manual sample preparation limits throughput.
This study demonstrates high-throughput MG screening using nMS for WEE1 binding to CRBN-DDB1,...
Bacterial pathogens often utilize post-translational modifications to manipulate host cells. Legionella pneumophila, the causative agent of Legionnaires' disease, secretes the enzyme AnkX, which is known to use cytidine diphosphate-choline to post-translationally modify the human small G-protein Rab1 with a phosphocholine moiety. Later during the infection, the Legionella enzyme Lem3 acts as a...
The Omnitrap-Orbitrap-Booster (OOB) mass spectrometry (MS) platform with enhanced high-resolution, fragmentation and data acquisition capabilities, was developed to advance top-down (TD) MS analysis of proteins. It integrates a multimodal tandem mass spectrometry (MS/MS) ion trap system (OmnitrapTM) offering a wide range of fragmentation methods, a high-resolution Orbitrap Fourier transform...
X-ray activation and dissociation of proteins and their non-covalent assemblies may elucidate structural and functional details complementary to established top-down mass-spectrometry techniques. This is attributed to the rapid, site-specific ionization of atoms within biomolecules. Our research group conducted proof-of-concept experiments exploring X-ray activation of samples with masses...
Trapped ion mobility spectrometry (TIMS) spatially separates ions when suspended in a high-pressure region between a retarding electric field gradient and constant gas flow from the atmospheric pressure inlet of the mass spectrometer. TIMS allows high resolution separation of ions/isomers/conformers with varying duty cycles in a relatively small device (<10cm). TIMS is also particularly well...
Abnormal protein phosphorylation is a fundamental trigger in the pathogenesis of Alzheimer’s Disease, leading to the formation of neurofibrillary tangles. Thus, molecular determination of the critical factors in controlling phosphorylation is in high demand. Pin1, a cis-trans prolyl isomerase has recently been implicated in Alzheimer’s Disease progression. Moreover, Pin1 specifically targets...
Monoclonal antibodies (mAbs) have revolutionized biotherapeutics, offering effective treatments for various diseases. Antibody-drug conjugates (ADCs) combine mAbs’ specificity with potent cytotoxic drugs linked via Lys- or Cys-conjugation, targeting malignant cells. However, heterogeneity in payload attachment challenges ADCs' safety and efficacy. We employed a middle-down (MD) mass...
