Related Experiment Video
Updated: Feb 12, 2026
Combining Wet and Dry Lab Techniques to Guide the Crystallization of Large Coiled-coil Containing Proteins
Published on: January 6, 2017
Engineered Coiled-Coil Protein for Delivery of Inverse Agonist for Osteoarthritis
Liming Yin1, Albert S Agustinus1, Carlo Yuvienco1
1Department of Chemical and Biomolecular Engineering , NYU Tandon School of Engineering , Brooklyn , New York 11201 , United States.
Abstract:
Osteoarthritis (OA) results from degenerative and abnormal function of joints, with localized biochemistry playing a critical role in its onset and progression. As high levels of all- trans retinoic acid (ATRA) in synovial fluid have been identified as a contributive factor to OA, the synthesis of de novo antagonists for retinoic acid receptors (RARs) has been exploited to interrupt the mechanism of ATRA action. BMS493, a pan-RAR inverse agonist, has been reported as an effective inhibitor of ATRA signaling pathway; however, it is unstable and rapidly degrades under physiological conditions. We employed an engineered cartilage oligomeric matrix protein coiled-coil (CccS) protein for the encapsulation, protection, and delivery of BMS493. In this study, we determine the binding affinity of CccS to BMS493 and the stimulator, ATRA, via competitive binding assay, in which ATRA exhibits approximately 5-fold superior association with CccS than BMS493. Interrogation of the structure of CccS indicates that ATRA causes about 10% loss in helicity, while BMS493 did not impact the structure. Furthermore, CccS self-assembles into nanofibers when bound to BMS493 or ATRA as expected, displaying 11-15 nm in diameter. Treatment of human articular chondrocytes in vitro reveals that CccS·BMS493 demonstrates a marked improvement in efficacy in reducing the mRNA levels of matrix metalloproteinase-13 (MMP-13), one of the main proteases responsible for the degradation of the extracellular cartilage matrix compared to BMS493 alone in the presence of ATRA, interleukin-1 beta (IL-1β), or IL-1 β together with ATRA. These results support the feasibility of utilizing coiled-coil proteins as drug delivery vehicles for compounds of relatively limited bioavailability for the potential treatment of OA.
Insights
Engineered coiled-coil proteins effectively deliver BMS493, an osteoarthritis drug, improving its efficacy and stability. This novel drug delivery system shows promise for treating osteoarthritis by protecting the drug and enhancing its therapeutic action.
Area of Science:
- Biochemistry
- Biomaterials Science
- Drug Delivery Systems
Background:
- Osteoarthritis (OA) involves joint degeneration, with all-trans retinoic acid (ATRA) in synovial fluid contributing to its progression.
- Retinoic acid receptor (RAR) antagonists are investigated to counteract ATRA's effects, but existing inhibitors like BMS493 lack stability under physiological conditions.
Purpose of the Study:
- To develop a stable drug delivery system for BMS493 using an engineered cartilage oligomeric matrix protein coiled-coil (CccS) protein.
- To evaluate the binding affinity, structural impact, and therapeutic efficacy of CccS·BMS493 in an osteoarthritis model.
Main Methods:
- Competitive binding assays were used to determine the binding affinities of ATRA and BMS493 to CccS.
- Circular dichroism spectroscopy assessed the structural impact of ATRA and BMS493 binding on CccS.
- The self-assembly of CccS upon binding was analyzed, and the efficacy of CccS·BMS493 was tested on human articular chondrocytes in vitro.
Main Results:
- ATRA showed a 5-fold higher association with CccS than BMS493; ATRA binding caused a 10% loss in CccS helicity, while BMS493 did not.
- CccS self-assembled into 11-15 nm nanofibers when bound to either BMS493 or ATRA.
- CccS·BMS493 significantly enhanced the reduction of matrix metalloproteinase-13 (MMP-13) mRNA levels in chondrocytes compared to BMS493 alone, particularly under inflammatory conditions.
Conclusions:
- Engineered coiled-coil proteins (CccS) can serve as effective carriers for unstable drugs like BMS493, improving their bioavailability and therapeutic efficacy.
- The CccS·BMS493 system demonstrates potential for treating osteoarthritis by mitigating cartilage degradation.
- This study validates the use of coiled-coil proteins as versatile drug delivery vehicles for compounds with limited bioavailability.
Related Concept Videos
02:04Protein-protein Interfaces
02:21Globular and Fibrous Proteins
Globular proteins are also known as spheroproteins and typically are approximately round in shape. They contain a mix of amino acid types and contain differing sequences in their primary structures. Globular proteins have many different functions, such as enzymes, cellular messengers, and molecular transporters. These roles often require the proteins to be...
00:49What is Genetic Engineering?
01:29Inverse Trigonometric Functions
01:25Inverse Hyperbolic Functions and Their Derivatives
01:30Adrenergic Agonists: Therapeutic Uses
Emergency and Intensive Care Unit (ICU) applications: Pressor agents increase blood pressure, heart rate, and contractility in shock and organ failure situations. Dopamine can induce vasodilation and stimulate adrenoceptors. Endogenous catecholamines are effective in treating cardiogenic shock. α2-agonists like clonidine can reverse anesthesia-induced hypertension.
Allergies and...