β2-mikroglobulino gryninimas ir skysčio-skysčio fazių atsiskyrimo tyrimai /

β2-microglobulin is an important component of the major histocompatibility complex class I (MHC-I) molecule, which is found on almost all nucleated cells. Its main role in the immune system is the presentation of peptide antigens to T cells. In addition to its physiological functions, β2M is clinically important because it can cause diseases called amyloidoses. The protein is eliminated by the kidneys and, in the event of kidney dysfunction, the accumulation of the protein in the human body promotes the formation of amyloid fibrils and can lead to dialysis-related amyloidosis. A rapidly growing area of research is liquid-liquid phase separation (LLPS), the process by which proteins and other biomolecules separate from the bulk solution to form dense, membraneless condensates. Although dozens of proteins have been identified to form condensates, there is currently no information on whether β2M can undergo LLPS. In the course of this work, the purification process of the protein was optimised. It was found that the highest yield of the protein is obtained by denaturation with guanidine hydrochloride and purification of the protein by ion-exchange chromatography with a Q-Sepharose sorbent. An investigation of the effect of environmental conditions (protein concentration, ionic strength of the solution, denaturant concentration and pH) on β2M LLPS showed that pH is the most important factor in this process. In the pH range of 4 to 5, β2M was able to form protein condensates and aggregates. This finding opens new avenues for understanding the behavior of β2M under different physiological and pathological conditions, potentially linking its phase separation properties to its role in disease mechanisms.

Grantor dc:publisher

Institutional Repository of Vilnius University

Year dc:date

2024

Author dc:creator

• Miknaitė, Justina,

Statement dc:rights

• info:eu-repo/semantics/openAccess

Language dc:language

lit

Repository record dc:identifier

https://repository.vu.lt/VU:ELABAETD210639988&prefLang=en_US

OAI identifier oai:identifier

oai:vu.lt:elaba:210639988