Physics Commons^™

Association Of Alpha-Crystallin With Human Cortical And Nuclear Lens Lipid Membrane Increases With The Grade Of Cortical And Nuclear Cataract, Preston Hazen, Geraline Trossi-Torres, Raju Timsina, Nawal K. Khadka, Laxman Mainali

Physics Faculty Publications and Presentations

Eye lens α-crystallin has been shown to become increasingly membrane-bound with age and cataract formation; however, to our knowledge, no studies have investigated the membrane interactions of α-crystallin throughout the development of cataracts in separated cortical membrane (CM) and nuclear membrane (NM) from single human lenses. In this study, four pairs of human lenses from age-matched male and female donors and one pair of male lenses ranging in age from 64 to 73 years old (yo) were obtained to investigate the interactions of α-crystallin with the NM and CM throughout the progression of cortical cataract (CC) and nuclear cataract (NC) …

Cholesterol Content Regulates The Interaction Of Αa-, Αb-, And Α-Crystallin With The Model Of Human Lens-Lipid Membranes, Raju Timsina, Preston Hazen, Geraline Trossi-Torres, Nawal K. Khadka, Navdeep Kalkat, Laxman Mainali

Physics Faculty Publications and Presentations

α-Crystallin (αABc) is a major protein comprised of αA-crystallin (αAc) and αB-crystallin (αBc) that is found in the human eye lens and works as a molecular chaperone by preventing the aggregation of proteins and providing tolerance to stress. However, with age and cataract formation, the concentration of αABc in the eye lens cytoplasm decreases, with a corresponding increase in the membrane-bound αABc. This study uses the electron paramagnetic resonance (EPR) spin-labeling method to investigate the role of cholesterol (Chol) and Chol bilayer domains (CBDs) in the binding of αAc, αBc, and αABc to the Chol/model of human lens-lipid (Chol/MHLL) membranes. …

Binding Of Alpha-Crystallin To Cortical And Nuclear Lens Lipid Membranes Derived From A Single Lens, Raju Timsina, Samantha Wellisch, Dieter Haemmerle, Laxman Mainali

Physics Faculty Publications and Presentations

Several studies reported that α-crystallin concentrations in the eye lens cytoplasm decrease with a corresponding increase in membrane-bound α-crystallin with age and cataracts. The influence of the lipid and cholesterol composition difference between cortical membrane (CM) and nuclear membrane (NM) on α-crystallin binding to membranes is still unclear. This study uses the electron paramagnetic resonance (EPR) spin-labeling method to investigate the α-crystallin binding to bovine CM and NM derived from the total lipids extracted from a single lens. Compared to CMs, NMs have a higher percentage of membrane surface occupied by α-crystallin and binding affinity, correlating with less mobility and …

Alpha-Crystallin Association With The Model Of Human And Animal Eye Lens-Lipid Membranes Is Modulated By Surface Hydrophobicity Of Membranes, Raju Timsina, Geraline Trossi-Torres, Jackson Thieme, Matthew O'Dell, Nawal K. Khadka, Laxman Mainali

Physics Faculty Publications and Presentations

Purpose

This research aims to probe the interaction of α-crystallin with a model of human, porcine, and mouse lens-lipid membranes.

Methods

Cholesterol/model of human lens-lipid (Chol/MHLL), cholesterol/model of porcine lens-lipid (Chol/MPLL), and cholesterol/model of mouse lens-lipid (Chol/MMLL) membranes with 0–60 mol% Chol were prepared using the rapid solvent exchange method and probe-tip sonication. The hydrophobicity near the surface of model lens-lipid membranes and α-crystallin association with these membranes were investigated using the electron paramagnetic resonance spin-labeling approach.

Results

With increased Chol content, the hydrophobicity near the surface of Chol/MHLL, Chol/MPLL, and Chol/MMLL membranes, the maximum percentage of membrane surface occupied …

Alpha-Crystallin-Membrane Association Modulated By Phospholipid Acyl Chain Length And Degree Of Unsaturation, Geraline Trossi-Torres, Raju Timsina, Laxman Mainali

Physics Faculty Publications and Presentations

α-crystallin-membrane association increases with age and cataracts, with the primary association site of α-crystallin being phospholipids. However, it is unclear if phospholipids’ acyl chain length and degree of unsaturation influence α-crystallin association. We used the electron paramagnetic resonance approach to investigate the association of α-crystallin with phosphatidylcholine (PC) membranes of different acyl chain lengths and degrees of unsaturation and with and without cholesterol (Chol). The association constant (K_a) of α-crystallin follows the trends, i.e., K_a (14:0–14:0 PC) > K_a (18:0–18:1 PC) > K_a (18:1–18:1 PC) ≈ K_a (16:0–20:4 PC) where the presence of Chol decreases K …

Association Of Alpha-Crystallin With Fiber Cell Plasma Membrane Of The Eye Lens Accompanied By Light Scattering And Cataract Formation, Raju Timsina, Laxman Mainali

Physics Faculty Publications and Presentations

α-crystallin is a major protein found in the mammalian eye lens that works as a molecular chaperone by preventing the aggregation of proteins and providing tolerance to stress in the eye lens. These functions of α-crystallin are significant for maintaining lens transparency. However, with age and cataract formation, the concentration of α-crystallin in the eye lens cytoplasm decreases with a corresponding increase in the membrane-bound α-crystallin, accompanied by increased light scattering. The purpose of this review is to summarize previous and recent findings of the role of the: (1) lens membrane components, i.e., the major phospholipids (PLs) and sphingolipids, cholesterol …

Cholesterol And Cholesterol Bilayer Domains Inhibit Binding Of Alpha-Crystallin To The Membranes Made Of The Major Phospholipids Of Eye Lens Fiber Cell Plasma Membranes, Raju Timsina, Geraline Trossi-Torres, Matthew O'Dell, Nawal K. Khadka, Laxman Mainali

Physics Faculty Publications and Presentations

The concentration of α-crystallin decreases in the eye lens cytoplasm, with a corresponding increase in membrane-bound α-crystallin during cataract formation. The eye lens’s fiber cell plasma membrane consists of extremely high cholesterol (Chol) content, forming cholesterol bilayer domains (CBDs) within the membrane. The role of high Chol content in the lens membrane is unclear. Here, we applied the continuous-wave electron paramagnetic resonance spin-labeling method to probe the role of Chol and CBDs on α-crystallin binding to membranes made of four major phospholipids (PLs) of the eye lens, i.e., phosphatidylcholine (PC), sphingomyelin (SM), phosphatidylserine (PS), and phosphatidylethanolamine (PE). Small unilamellar vesicles …

Interaction Of Alpha-Crystallin With Phospholipid Membranes, Laxman Mainali, William J. O'Brien, Raju Timsina

Physics Faculty Publications and Presentations

Purpose/Aim: The amount of membrane-bound α-crystallin increases significantly with age and cataract formation, accompanied by a corresponding decline in the level of α-crystallin in the lens cytoplasm. The purpose of this research is to evaluate the binding affinity of α-crystallin to the phospholipid membranes as well as the physical properties of the membranes after α-crystallin binding.

Materials and Methods: The continuous wave and saturation recovery electron paramagnetic resonance (EPR) methods were used to obtain the information about the binding affinity and the physical properties of the membrane. In this approach, the cholesterol analogue spin label CSL was incorporated in the …

Interaction Of Alpha-Crystallin With Four Major Phospholipids Of Eye Lens Membranes, Raju Timsina, Nawal K. Khadka, David Maldonado, Laxman Mainali

Physics Faculty Publications and Presentations

It is well-studied that the significant factor in cataract formation is the association of α-crystallin, a major eye lens protein, with the fiber cell plasma membrane of the eye lens. The fiber cell plasma membrane of the eye lens consists of four major phospholipids (PLs), i.e., phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and sphingomyelin (SM). Despite several attempts to study the interaction of α-crystallin with PLs of the eye lens membrane, the role of individual PL for the binding with α-crystallin is still unclear. We recently developed the electron paramagnetic resonance (EPR) spin-labeling method to study the binding of α-crystallin …