PXD041226

PXD041226 is an original dataset announced via ProteomeXchange.

Title	From Volcanos to the Bench: Advantages of Novel Hyperthermoacidic Archaeal Proteases for Proteomics Workflows
Description	Here we introduce novel hyperthermoacidic archaeal proteases (HTA-Proteases) isolated from organisms that thrive in nearly boiling acidic volcanic springs and investigate their use for bottom-up proteomic experiments. We find that HTA-Proteases have novel cleavage specificities, show no autolysis, function in dilute formic acid, and store at ambient temperature for years. HTA-Proteases function optimally at 70-90°C and pH of 2-4 with rapid digestion kinetics. The extreme reaction conditions actively denature sample proteins, obviate the use of chaotropes, and allow for a one-step/five-minute sample preparation protocol without sample manipulation, dilution, or additional cleanup. We find that brief one-step HTA-Protease protocols significantly increase proteome and protein sequence coverage with datasets orthogonal to trypsin. Importantly, HTA-Protease digests markedly increase coverage and identifications for ribonucleoproteins, histones, and mitochondrial membrane proteins as compared to tryptic digests. In addition to increased coverage in these classes, HTA-Proteases and simplified one-step protocols are expected to reduce technical variability and advance the fields of clinical and high-throughput proteomics. This work reveals significant utility of heretofore unavailable HTA-Proteases for proteomic workflows. We discuss some of the potential for these remarkable enzymes to empower new proteomics methods, approaches, and biological insights.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:24:04.267.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Maxwell McCabe
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Bos taurus (Bovine); NCBI TaxID: 9913; scientific name: Sus scrofa domesticus (domestic pig); NCBI TaxID: 9825; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monomethylated residue; acetylated residue; monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	LTQ XL; Q Exactive HF; LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2023-03-30 14:39:21	ID requested
1	2023-10-18 10:38:05	announced
⏵ 2	2023-11-14 08:24:04	announced	2023-11-14: Updated project metadata.

Dataset with its publication pending

submitter keyword: heat, hydrolysis,Archaea, protease, ultrastable enzymes, hyperthermophile, sample preparation, acid, digestion

Kirk Hansen
contact affiliation	Department of Biochemistry and Molecular Genetics, University of Colorado Denver, Aurora, CO, 80045, USA
contact email	kirk.hansen@cuanschutz.edu
lab head
Maxwell McCabe
contact affiliation	University of Colorado
contact email	maxwell.mccabe@cuanschutz.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD041226
     1. Label: PRIDE project
     2. Name: From Volcanos to the Bench: Advantages of Novel Hyperthermoacidic Archaeal Proteases for Proteomics Workflows