TY - JOUR
T1 - Reconstitution of LDL with lipophilic fluorescein derivatives
T2 - Quantitative analysis of the receptor activity of human lymphocytes
AU - Krieger, Monty
AU - Ho, Y. K.
AU - Falck, J R
N1 - Funding Information:
We are grateful to Professor Julian Peterson and Mr. Peter Roome who provided valuable assistance in recording the absorption spectra and to Dr. David Bilheimer for his cooperation in obtaining subjects for this work. This work was supported by USPHS grants PO 1-HL-20948 and HL05657 from the National Institutes of Health. It was performed in the laboratories of Michael Brown and Joseph Goldstein to whom we are grateful for advise, support and a critical evaluation of this manuscript. Dr. Krieger's current address is the Biology Department and the Whitaker College of Health Sciences, Technology and Management, Massachusetts Institute of Technology, Cambridge, MA 02139.
PY - 1983
Y1 - 1983
N2 - Low density lipoprotein (LDL), the major cholesterol transport protein in human plasma, consists of an apolar core of cholesteryl esters surrounded by a polar shell containing phospho-lipids, unesterified cholesterol and protein. In the current paper we report the absorption and fluorescence spectra of members of a new class of lipophilic fluorescein derivatives which were designed to be reconstituted into the core of LDL in place of the native cholesteryl esters. One of these derivates, cholesteryl 12-0-[methyl 3-0-methyl-5′6′carboxyfluorescein]ricinoleyl carbonate (MMC) was reconstituted into the core of LDL. The resultant fluorescent reconstituted LDL was used in conjunction with flow cytometry to quantify the LDL receptor activity of fresh blood lymphocytes derived from normal individuals and from patients with the heterozygous and homozygous forms of familial hypercholesterolemia (FH). The LDL receptor activities of the heterozygous and homozygous FH lymphocytes were approximately 37% and 1% of normal, respectively. LDL reconstituted with these lipophilic fluorescein derivatives will be valuable in studying LDL metabolism and may be useful for the diagnosis of FH.
AB - Low density lipoprotein (LDL), the major cholesterol transport protein in human plasma, consists of an apolar core of cholesteryl esters surrounded by a polar shell containing phospho-lipids, unesterified cholesterol and protein. In the current paper we report the absorption and fluorescence spectra of members of a new class of lipophilic fluorescein derivatives which were designed to be reconstituted into the core of LDL in place of the native cholesteryl esters. One of these derivates, cholesteryl 12-0-[methyl 3-0-methyl-5′6′carboxyfluorescein]ricinoleyl carbonate (MMC) was reconstituted into the core of LDL. The resultant fluorescent reconstituted LDL was used in conjunction with flow cytometry to quantify the LDL receptor activity of fresh blood lymphocytes derived from normal individuals and from patients with the heterozygous and homozygous forms of familial hypercholesterolemia (FH). The LDL receptor activities of the heterozygous and homozygous FH lymphocytes were approximately 37% and 1% of normal, respectively. LDL reconstituted with these lipophilic fluorescein derivatives will be valuable in studying LDL metabolism and may be useful for the diagnosis of FH.
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U2 - 10.3109/10799898309041846
DO - 10.3109/10799898309041846
M3 - Article
C2 - 6312035
AN - SCOPUS:0020690404
SN - 1079-9893
VL - 3
SP - 361
EP - 375
JO - Journal of Receptors and Signal Transduction
JF - Journal of Receptors and Signal Transduction
IS - 3
ER -