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

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 (K-7 H[Nb6O19 ] .13H (2) O, Nb O-6(19) ) 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 Nb 6 O 19 /MIL-125(Ti) can promote the selective adsorption of Cyt-C due to the synergistic effect of electrostatic and hydrogenbond interactions. At pH = 10.0 (Britton-Robinson buffer), the adsorption efficiency of 30 0 mu L 10 0 mu g .mL (-1) Cyt-C onto 1.0 mg Nb (6) O (19) /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 Nb (6) O (19) /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.

Automated summary

In this study, a high efficient and selective pretreatment method for Cytochrome C (Cyt-C) in complex protein samples was developed using a polyniobate-modified MOF material. The material exhibited selective adsorption of Cyt-C due to the synergistic effect of electrostatic and hydrogen bond interactions. The adsorption and separation efficiency of Cyt-C was validated through SDS-PAGE and LC-MS/MS analysis.