New Mass-Spectrometry-Compatible Degradable Surfactant for Tissue Proteomics

Abstract

Tissue proteomics is increasingly recognized for its role in biomarker discovery and disease mechanism investigation. However, protein solubility remains a significant challenge in mass spectrometry (MS)-based tissue proteomics. Conventional surfactants such as sodium dodecyl sulfate (SDS), the preferred surfactant for protein solubilization, are not compatible with MS. Herein, we have screened a library of surfactant-like compounds and discovered an MS-compatible degradable surfactant (MaSDeS) for tissue proteomics that solubilizes all categories of proteins with performance comparable to SDS. The use of MaSDeS in the tissue extraction significantly improves the total number of protein identifications from commonly used tissues, including tissue from the heart, liver, and lung. Notably, MaSDeS significantly enriches membrane proteins, which are often under-represented in proteomics studies. The acid degradable nature of MaSDeS makes it amenable for high-throughput mass spectrometry-based proteomics. In addition, the thermostability of MaSDeS allows for its use in experiments requiring high temperature to facilitate protein extraction and solubilization. Furthermore, we have shown that MaSDeS outperforms the other MS-compatible surfactants in terms of overall protein solubility and the total number of identified proteins in tissue proteomics. Thus, the use of MaSDeS will greatly advance tissue proteomics and realize its potential in basic biomedical and clinical research. MaSDeS could be utilized in a variety of proteomics studies as well as general biochemical and biological experiments that employ surfactants for protein solubilization.

Less is More: Membrane Protein Digestion Beyond Urea–Trypsin Solution for Next-level Proteomics

“…However, these detergents vary solubilization power with proteins (some instantly precipitate proteins such as cytochrome c oxidase, CcO), require chemical cleavage post-digestion-difficult for fast full automation, and some cleaved products contain amines that interfere with TMT. A latest acid-cleavable analog of SDS, anionic MaSDeS, showed SDS-like strong solubilization power on tissues, though it is severely denaturing and forms amine products (43,44). Amphi-polymers (amphipols) can support protein extraction and digestion, but most aggregate below pH 3-4 and require removal pre-HPLC by precipitation and centrifugation (45), interrupting automatable workflow.…”

A Family of Photolabile Nitroveratryl-Based Surfactants That Self-Assemble into Photodegradable Supramolecular Structures

“…53 These surfactants are also potentially useful as removable templates for the synthesis of meso- or microporous materials 4 or as removable surface-acting agents for extracting hydrophobic proteins. 11 These represent a new type of highly versatile photolabile surfactants that could have a wide range of potential applications.…”

“…46 High-resolution Fourier transformed ion cyclotron resonance (FT-ICR) mass spectra before and after UV irradiation were obtained on a 7 T LTQ/FT (Thermo Scientific, Waltham, MA), similarly as described in our previous publication. 11 DLS was carried out using a Malvern Zetasizer Nano. All DLS measurements of individual samples were performed three times after 120 sec equilibration.…”

“…1-5 They are also desirable in organic synthesis because the nano- and micro-structures formed by surfactants can be used as a reaction vessel to allow for compartmentalization of the reactions. 6-10 Furthermore, they are commonly used in many biological applications as surface-acting reagents for extracting and solubilizing proteins from cell or tissue lysates, 11-13 or as drug delivery containers for water insoluble drugs or gene delivery containers. 14-18 Through the addition of stimuli sensitive moieties into the molecular structure of the surfactant, a responsive surfactant can be created that allows for the external control of its properties in a predicted manner.…”

“…25 Moreover, because a small amount of ionic surfactant molecules can suppress the mass spectrometry (MS) signals, stimuli-labile surfactants are especially attractive because it allows for the extraction of proteins from cell or tissue that can be use for further MS analysis. 11 Therefore, the development of stimuli-labile surfactant molecules is highly desirable for applications in various chemical and biological fields. These stimuli can range from a change in the solution pH, 26-28 ionic strength, redox environment, 29 to light 20, 22, 23, 30, 31 and temperature.…”