Isolation of Cytochrome C for Proteomics with Lindqvist-type Polyiodate Modified Metal Organic Framework

Xi Wu; Quanxing Mao; Yangguang Hao; Junna Yang; Xiaonan Zhang; Zixin Chi; Guangyan Liu; Mengmeng Wang; Qing Chen; Xuwei Chen

doi:10.1016/j.chroma.2023.463869

Isolation of Cytochrome C for Proteomics with Lindqvist-type Polyiodate Modified Metal Organic Framework

J Chromatogr A. 2023 Mar 29:1693:463869. doi: 10.1016/j.chroma.2023.463869. Epub 2023 Feb 19.

Authors

Xi Wu¹, Quanxing Mao¹, Yangguang Hao², Junna Yang¹, Xiaonan Zhang², Zixin Chi², Guangyan Liu², Mengmeng Wang³, Qing Chen⁴, Xuwei Chen⁵

Affiliations

¹ College of Chemistry, Liaoning University, Shenyang, 110036, People's Republic of China.
² Institute of Translational Medicine, Department of Pharmacy, Shenyang Medical College, Shenyang, 110034, People's Republic of China.
³ Institute of Translational Medicine, Department of Pharmacy, Shenyang Medical College, Shenyang, 110034, People's Republic of China;. Electronic address: 1450989834@qq.com.
⁴ Institute of Translational Medicine, Department of Pharmacy, Shenyang Medical College, Shenyang, 110034, People's Republic of China;. Electronic address: chenqing.0906@163.com.
⁵ Research Center for Analytical Sciences, Department of Chemistry, College of Sciences, Northeastern University, Shenyang 110819, People's Republic of China.

PMID: 36822038
DOI: 10.1016/j.chroma.2023.463869

Abstract

Separation and purification of Cytochrome C (Cyt-C) is important for proteomic. High efficient and selective pretreatment method for Cyt-C in real samples are always needed. Herein, polyniobate (K₇H[Nb₆O₁₉]·13H₂O, Nb₆O₁₉) is modified on a metal-organic framework MIL-125(Ti) through intermolecular hydrogen bonds and an aqueous-stable composite Nb₆O₁₉/MIL-125(Ti) is successfully prepared to pretreat complex protein sample. Protein adsorption studies have shown that Nb₆O₁₉/MIL-125(Ti) can promote the selective adsorption of Cyt-C due to the synergistic effect of electrostatic and hydrogen-bond interactions. At pH=10.0 (Britton-Robinson buffer), the adsorption efficiency of 300 μL 100 μg·mL^-1 Cyt-C onto 1.0 mg Nb₆O₁₉/MIL-125(Ti) can reach 99.5%. The adsorption behavior of Cyt-C fits well with the Langmuir adsorption model, corresponding to a maximum theoretical adsorption capacity of 168.35 mg·g^-1. Using 3 mol·L^-1 NaCl as the eluent, a high elution efficiency of 92.19% is obtained. In addition, the results of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis confirm that Nb₆O₁₉/MIL-125(Ti) efficiently adsorbed Cyt-C from scrofa heart extraction. LC-MS/MS spectrometry results show that the purification of Cyt-C reduces the abundance from the 12th to the 154th place after Nb₆O₁₉/MIL-125(Ti) treatment. Moreover, low abundant proteins, e.g., Superoxide dismutase 1, IF rod domain-containing protein and Ubiquitin-60S ribosomal protein L40 were considerably enriched. These outcomes confirm the practicability of Nb₆O₁₉/MIL-125 (Ti) as a Cyt-C extractant has potential application value in scrofa heart proteomics.

Keywords: Cyt-C; Lindqvist-type polyniobate; heart proteomics; metal organic framework; separation and purification.

MeSH terms

Adsorption
Chromatography, Liquid
Cytochromes c / metabolism
Metal-Organic Frameworks* / chemistry
Proteomics
Tandem Mass Spectrometry

Substances

Metal-Organic Frameworks
Cytochromes c