<abstract xml:lang="en" xmlns="http://www.w3.org/1999/xhtml"><p>Although described some time ago, gamma-butyrobetaine esters and related compounds have not gained much attention from researchers, and their physiological function remains obscure. Formerly we detected GBB-esterase enzymatic activity in rat blood serum. The aim of the present work was to develop a protocol that would enable purification of the protein fraction enriched in GBB esterase activity from rat blood serum. Chromatography on DEAE Sepharose at pH 4.2 enabled to purify a protein fraction enriched in enzymatic activity, but represented by numerous polypeptides. Following separation of this fraction by means of chromatography on DEAE Sepharose at pH 6.5 or heparin Sepharose chromatography at pH 7.0 did not lead to significant decrease of polypeptide number. When the above fraction was further fractionated by means of DEAE Sepharose chromatography at pH 7.4 or Bio Gel P150 chromatography the enzymatic activity was lost. Combination of DEAE Sepharose at pH 4.2 and affinity chromatography with procainamide appears to be the most suitable approach.</p></abstract>

Although described some time ago, gamma-butyrobetaine esters and related compounds have not gained much attention from researchers, and their physiological function remains obscure. Formerly we detected GBB-esterase enzymatic activity in rat blood serum. The aim of the present work was to develop a protocol that would enable purification of the protein fraction enriched in GBB esterase activity from rat blood serum. Chromatography on DEAE Sepharose at pH 4.2 enabled to purify a protein fraction enriched in enzymatic activity, but represented by numerous polypeptides. Following separation of this fraction by means of chromatography on DEAE Sepharose at pH 6.5 or heparin Sepharose chromatography at pH 7.0 did not lead to significant decrease of polypeptide number. When the above fraction was further fractionated by means of DEAE Sepharose chromatography at pH 7.4 or Bio Gel P150 chromatography the enzymatic activity was lost. Combination of DEAE Sepharose at pH 4.2 and affinity chromatography with procainamide appears to be the most suitable approach.

Protocol for purification of a rat blood serum protein fraction enriched in gamma-butyrobetaine esterase activity

Department of Biochemistry and Biophysics, Vilnius University, M. K. Čiurlionio 21, Vilnius, LT-2009, LITHUANIA

Latvian Institute of Organic Synthesis, Aizkraukles 21, Rīga, LV1006, LATVIA

Proceedings of the Latvian Academy of Sciences. Section B. Natural, Exact, and Applied Sciences.

Although described some time ago, gamma-butyrobetaine esters and related compounds have not gained much attention from researchers, and their physiological...

Protocol for purification of a rat blood serum protein fraction enriched in gamma-butyrobetaine esterase activity

Pubblicato online: 06 nov 2009

Pagine: 93 - 99

DOI: https://doi.org/10.2478/v10046-009-0023-8

Parole chiave
gamma-buturobetaine esterase, gamma-butyrobetaine esters, blood serum, affinity chromatography, proacainamide

This content is open access.

Protocol for purification of a rat blood serum protein fraction enriched in gamma-butyrobetaine esterase activity

Pubblicato online: 06 nov 2009

Pagine: 93 - 99

DOI: https://doi.org/10.2478/v10046-009-0023-8

Parole chiavegamma-buturobetaine esterase, gamma-butyrobetaine esters, blood serum, affinity chromatography, proacainamide

This content is open access.

Parole chiave
gamma-buturobetaine esterase, gamma-butyrobetaine esters, blood serum, affinity chromatography, proacainamide