TY - JOUR
T1 - Isolation, characterization, and assay of lecithin-cholesterol acyltransferase
AU - Albers, John J.
AU - Chen, Ching Hong
AU - Lacko, Andras G.
N1 - Funding Information:
Research of J.J.A. and C.-H. Chen referred to in this chapter was supported by the Natural Heart, Lung, and Blood Institute (HL-30086), and that of A.G.L. by the National Institutes of Health (AG-03255), the American Heart Association (82-1043), and the Robert A. Welch Foundation (B-935).
Funding Information:
The authors wish to acknowledget heir colleagues who were responsible for much of the work presented in this chapter. These individualsi nclude Drs. Alan Cardin, Moti Kashyap, Judith A. K. Harmony, Kohji Shirai, S. Ranganathan, George Holdsworlh, A. Balasubra-manium, Norihiro Sasaki, Daniel Quinn, Nobuo Matsuoka, and J. David Johnson, and Ms. Janet Simons who has prepared the manuscriptf or publication. This research was supported by U.S. Public Health Service Grants P01-HL-22619 and R01-HL-23019. L.R.M. was supported by a Molecular and Cellular BiologyT raining Grant NIH HL-07527.
PY - 1986/1/1
Y1 - 1986/1/1
N2 - Lecithin–cholesterol acyltransferase (LCAT) is synthesized by the liver and secreted into the bloodstream, where it is believed to act principally on the surface of high-density lipoproteins (HDL). The first attempts to isolate LCAT yielded only partially purified preparations even though a variety of chromatographic techniques were used, including hydroxylapatite adsorption, salt precipitation, ultracentrifugation, and affinity chromatography. Lecithin–cholesterol acyltransferase has been one of the most difficult enzymes to characterize from the physical, chemical, and enzymological points of view. The painstakingly slow early development of an efficient purification procedure and the instability of the enzyme precluded rapid progress. For the assay of LCAT activity, a stable, efficient, and uniform substrate that can be added to the enzyme source under conditions such that the concentration of substrate is not rate limiting is required. This method permits the preparation of large amounts of stable, efficient homogeneous and well-defined substrate that is suitable for measuring the enzyme activity in plasma fractions.
AB - Lecithin–cholesterol acyltransferase (LCAT) is synthesized by the liver and secreted into the bloodstream, where it is believed to act principally on the surface of high-density lipoproteins (HDL). The first attempts to isolate LCAT yielded only partially purified preparations even though a variety of chromatographic techniques were used, including hydroxylapatite adsorption, salt precipitation, ultracentrifugation, and affinity chromatography. Lecithin–cholesterol acyltransferase has been one of the most difficult enzymes to characterize from the physical, chemical, and enzymological points of view. The painstakingly slow early development of an efficient purification procedure and the instability of the enzyme precluded rapid progress. For the assay of LCAT activity, a stable, efficient, and uniform substrate that can be added to the enzyme source under conditions such that the concentration of substrate is not rate limiting is required. This method permits the preparation of large amounts of stable, efficient homogeneous and well-defined substrate that is suitable for measuring the enzyme activity in plasma fractions.
UR - http://www.scopus.com/inward/record.url?scp=0022556169&partnerID=8YFLogxK
U2 - 10.1016/0076-6879(86)29103-X
DO - 10.1016/0076-6879(86)29103-X
M3 - Article
C2 - 3014276
AN - SCOPUS:0022556169
SN - 0076-6879
VL - 129
SP - 763
EP - 783
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -