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Proteomic Analysis of Ginkgolic Acid Targets in HCCLM3 Cells via DARTS-LC-MS/MS

ABSTRACT: This study aims to identify the potential protein targets of Ginkgolic Acid in human hepatocellular carcinoma HCCLM3 cells using the Drug Affinity Responsive Target Stability (DARTS) assay combined with LC-MS/MS proteomics. Whole cell lysates from Ginkgolic Acid-treated and vehicle control (DMSO) HCCLM3 cells were subjected to limited proteolysis with proteinase K. The digested protein samples were separated by SDS-PAGE, and differential protein bands were excised, digested in-gel, and analyzed by mass spectrometry. The raw data will be used to identify proteins whose stability is altered upon Ginkgolic Acid binding, providing insights into its anti-tumor mechanisms.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Huizhen Zou

LAB HEAD: shuguo zheng

PROVIDER: PXD073111 | Pride | 2026-03-30

REPOSITORIES: Pride

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Source:

			Action	DRS
	480_FPEP240062611-1A.msf	Msf
	480_FPEP240062611-1A.pdResult	Other
	480_FPEP240062611-1A.raw	Raw
	480_FPEP240062612-1A.msf	Msf
	480_FPEP240062612-1A.pdResult	Other

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Project description:Diabetic wounds require continuous and coordinated modulation of the microenvironment concurrent with tissue regeneration, yet this remains a significant challenge. As a proof of concept, we herein propose to use dimeric copper peptide for diabetic wound treatment. The dimeric copper peptide (D-CuP) was first synthesized and then incorporated into a reactive oxygen species (ROS)-responsive hydrogel matrix to improve therapeutic compliance, culminating in the formulation of G/D-CuP. Compared to monomer copper peptide (CuP), a wound healing agent, D-CuP exhibits multivalency, enhanced biologic stability against proteases and the broad biological activities, such as anti-inflammatory and antioxidative properties, while promoting angiogenesis and fibroblast proliferation and migration. Meanwhile, the hydrogel matrix, exhibiting excellent ROS-scavenging capabilities, has been engineered to an intelligent drug reservoir for wound-responsive release of D-CuP at the wound site while simultaneously attenuating inflammatory responses. Ultimately, the G/D-CuP group demonstrated superior therapeutic efficacy, achieving in 97.2% closure of infected wounds.In this study, we present a proof-of-concept study to optimize the treatment of diabetic wounds. This method integrates a dimeric copper peptide hydrogel (G/D-CuP) to address several critical factors: i) The design and synthesis of the dimeric copper peptide (D-CuP) feature a broad range of targets and enhanced biological activities, such as anti-inflammatory and antioxidative properties, while promoting angiogenesis and fibroblast proliferation and migration; ii) Utilizing the steric hindrance conferred by dimerization reduces protease access to the active site and significantly improving stability against proteases; iii) A ROS-responsive hydrogel matrix has been synthesized, exhibiting superior ROS-scavenging capabilities, functioning as an intelligent drug reservoir that enables the controlled release of D-CuP at the wound interface while simultaneously attenuating inflammatory responses in the microenvironment; iv) G/D-CuP possesses outstanding deformability and spreadability, allowing it to adapt to any wound shape; v) The synthesis and preparation process of G/D-CuP are straightforward and cost-effective, making it highly promising for clinical translation. This multifunctional treatment platform is anticipated to accommodate the complex and dynamic biological processes involved in diabetic wound healing, thereby significantly improving the overall efficacy of wound healing treatments.

Dataset Information

Proteomic Analysis of Ginkgolic Acid Targets in HCCLM3 Cells via DARTS-LC-MS/MS

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