Thermal Denaturation of Fresh Frozen Tissue Enhances Mass Spectrometry Detection of Peptides.

Autor:	Kruse ARS; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee 37212, United States., Judd AM; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37212, United States., Gutierrez DB; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37212, United States., Allen JL; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37212, United States., Dufresne M; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee 37212, United States., Farrow MA; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee 37212, United States., Powers AC; Department of Medicine, Division of Diabetes, Endocrinology, and Metabolism, Vanderbilt University School of Medicine, Nashville, Tennessee 37212, United States.; VA Tennessee Valley Healthcare System, Nashville, Tennessee 37212, United States., Norris JL; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee 37212, United States.; Bruker Daltonics, Billerica 01821, Massachusetts United States.; Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37212, United States., Caprioli RM; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Chemistry, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Medicine, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37212, United States., Spraggins JM; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee 37212, United States.; Department of Chemistry, Vanderbilt University, Nashville, Tennessee 37212, United States.
Jazyk:	angličtina
Zdroj:	Analytical chemistry [Anal Chem] 2024 Oct 22; Vol. 96 (42), pp. 16861-16870. Date of Electronic Publication: 2024 Oct 11.
DOI:	10.1021/acs.analchem.4c03625
Abstrakt:	Thermal denaturation (TD), known as antigen retrieval, heats tissue samples in a buffered solution to expose protein epitopes. Thermal denaturation of formalin-fixed paraffin-embedded samples enhances on-tissue tryptic digestion, increasing peptide detection using matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI IMS). We investigated the tissue-dependent effects of TD on peptide MALDI IMS and liquid chromatography-tandem mass spectrometry signal in unfixed, frozen human colon, ovary, and pancreas tissue. In a triplicate experiment using time-of-flight, orbitrap, and Fourier-transform ion cyclotron resonance mass spectrometry platforms, we found that TD had a tissue-dependent effect on peptide signal, resulting in a (22.5%) improvement in peptide detection from the colon, a (73.3%) improvement in ovary tissue, and a (96.6%) improvement in pancreas tissue. Biochemical analysis of identified peptides shows that TD facilitates identification of hydrophobic peptides.
Databáze:	MEDLINE
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