Purification of a membrane-associated serine esterase from murine cytotoxic T lymphocytes by a single reverse phase column
ORCiD
David M. Ojcius: 0000-0003-1461-4495
Department
Biomedical Sciences
Document Type
Article
Publication Title
Protein Expression and Purification
ISSN
1046-5928
Volume
1
Issue
1
DOI
10.1016/1046-5928(90)90049-5
First Page
77
Last Page
80
Publication Date
9-1-1990
Abstract
The plasma and organelle membranes of a cytotoxic T lymphocyte line, CTLL-R8, were isolated by subcellular fractionation. After dissolving in detergent-containing buffer, the membrane proteins were isolated by high-performance liquid chromatography on a single reverse-phase column. The serine esterase activity in the fractions was detected by measuring hydrolysis of the ester compound Nα-benzyloxycarbonyl-l-lysine thiobenzyl ester. A major band was revealed in the fraction with highest serine esterase activity. Under sodium dodecyl sulfate-polyacrylamide gel electrophoresis, this band assumes a molecular weight of about 30 kDa. The amino-terminal sequence of the protein was analyzed and shows 100% identity with that of MCSP-3/granzyme F, a soluble serine esterase previously identified in the cytoplasmic granules of cytotoxic T lymphocytes. Modifications of this reverse-phase column method would thus represent a simple, convenient strategy for obtaining high yields of all the lymphocyte surface proteases, which could then be further characterized for function.
Recommended Citation
Jiang, S.,
Hasselkus-Light, C. S.,
Ojcius, D. M.,
&
Young, J. D.
(1990).
Purification of a membrane-associated serine esterase from murine cytotoxic T lymphocytes by a single reverse phase column.
Protein Expression and Purification, 1(1), 77–80.
DOI: 10.1016/1046-5928(90)90049-5
https://scholarlycommons.pacific.edu/dugoni-facarticles/185
