Topics

ExteNDing proteome coverage with legumain as highly specific digestion protease.

07:00 EST 17th January 2020 | BioPortfolio

Summary of "ExteNDing proteome coverage with legumain as highly specific digestion protease."

Bottom-up mass spectrometry-based proteomics utilizes proteolytic enzymes with well character-ized specificities to generate peptides amenable for identification by high throughput tandem mass spectrometry. Trypsin, which cuts specifically after the basic residues lysine and arginine, is the predominant enzyme used for proteome digestion, although proteases with alternative specificity are required to detect sequences that are not accessible after tryptic digest. Here, we show that the hu-man cysteine protease legumain exhibits strict substrate specificity for cleavage after asparagine and aspartic acid residues during in-solution digestions of proteomes extracted from E.coli, mouse em-bryonic fibroblast cell cultures and Arabidopsis thaliana leaves. Generating peptides highly com-plementary in sequence, yet similar in their biophysical properties, legumain enabled complemen-tary proteome and protein sequence coverage as compared to trypsin or GluC. Importantly, legu-main further enabled the identification and enrichment of protein N-termini not accessible in GluC- or trypsin-digested samples. Legumain cannot cleave after glycosylated Asn residues, which ena-bled robust identification and orthogonal validation of N-glycosylation sites based on alternating sequential sample treatment with legumain and PNGaseF and vice versa. Taken together, we demonstrate that legumain is a practical, efficient protease for extending the proteome and sequence coverage achieved with trypsin, with unique possibilities for the characterization of post-translational modification sites.

Affiliation

Journal Details

This article was published in the following journal.

Name: Analytical chemistry
ISSN: 1520-6882
Pages:

Links

DeepDyve research library

PubMed Articles [14274 Associated PubMed Articles listed on BioPortfolio]

Increased Legumain/Smad3 expression in atherosclerotic plaque of rat thoracic aorta.

The purpose of this study was to investigate the role of legumain in the formation and stability of atherosclerotic plaque, as well as to explore the association between legumain with Smad3 pathway in...

N-Terminomics/TAILS Profiling of Macrophages after Chemical Inhibition of Legumain.

Legumain (asparaginyl endopeptidase) is the only protease with a preference for cleavage after asparagine residues. Increased legumain activity is a hallmark of inflammation, neurodegenerative disease...

Improving the Sequence Coverage of Integral Membrane Proteins during Hydrogen/Deuterium Exchange Mass Spectrometry Experiments.

Insight into the structure-function relationship of membrane proteins is important to understand basic cell function and inform drug development, as these are common targets for drugs. Hydrogen/deuter...

Improved RNA modification mapping of cellular non-coding RNAs using C- and U-specific RNases.

Locating ribonucleoside modifications within an RNA sequence requires digestion of the RNA into oligoribonucleotides of amenable size for subsequent analysis by LC-MS (liquid chromatography-mass spect...

Legumain acts on neuroinflammatory to affect CUS-induced cognitive impairment.

Cognitive impairment has been widely recognized as a central feature of depression. Legumain, a lysosomal cysteine protease, plays an important role in cancer, atherosclerosis, inflammation and other ...

Clinical Trials [3989 Associated Clinical Trials listed on BioPortfolio]

Starches Digestion After Obesity Surgery.

Despite an impressive capacity to induce diabetes remissions, the gastric bypass surgery has been associated with the onset of hyperglycemic peaks, which are very intensive and transient, ...

Proteome Remodelling

The purpose of the project is to investigate the proteome signature dynamics and remodelling in human skeletal muscle induced by exercise training

Effect of Xen Implantation on the Aqueous Humor Proteome

The purpose of this study is to determine how the aqueous humor proteome changes after implantation of a Xen microstent for treatment of glaucoma.

Gastric Emptying of Rice With Different Starch Properties

Glycemic carbohydrates are associated with metabolic disturbances, such as type II diabetes, due to rapid digestion of starch into glucose. The specific properties of starch within these f...

Alterations in the Plasma Proteome of Early-Onset Severe Preeclampsia

The hypothesis of this study is that many plasma proteins are altered in concentration and structure in preeclampsia and the elucidation of these alterations will add to the poorly underst...

Medical and Biotech [MESH] Definitions

Public Law 104-91, enacted in 1996, is designed to protect health insurance coverage for workers and their families when they change or lose their jobs. HIPAA has separate provisions for the large and small group markets, and the individual market. HIPAA amends the Employee Retirement Income Security Act (ERISA), the Public Health Service Act, and the Internal Revenue Code to provide improved portability and continuity of health insurance coverage, extending earlier provisions under the Consolidated Omnibus Budget Reconciliation Act of 1985 ("COBRA").

Techniques for measuring specific nucleic acid interaction with another nucleic acid or with a protein by digestion of the non-interacting nucleic acid by various nucleases. After all non-interacting regions are eliminated by nuclease digestion, the protected nucleic acid that remains is analyzed. DNA FOOTPRINTING utilizes this technique to analyze the DNA contact sites of DNA-BINDING PROTEINS.

Health insurance coverage for all persons in a state or country, rather than for some subset of the population. It may extend to the unemployed as well as to the employed; to aliens as well as to citizens; for pre-existing conditions as well as for current illnesses; for mental as well as for physical conditions.

Insurance coverage of a physical or mental health condition, disability, or illness existing before health insurance is applied for or enrolled in.

Insurance providing coverage of medical, surgical, or hospital care in general or for which there is no specific heading.

Quick Search


DeepDyve research library

Relevant Topics

Bioinformatics
Bioinformatics is the application of computer software and hardware to the management of biological data to create useful information. Computers are used to gather, store, analyze and integrate biological and genetic information which can then be applied...

Enzymes
Enzymes are proteins that catalyze (i.e., increase the rates of) chemical reactions. In enzymatic reactions, the molecules at the beginning of the process, called substrates, are converted into different molecules, called products. Almost all chemical re...


Searches Linking to this Article