Partial purification of the 5-hydroxytryptophan-reuptake system from human blood platelets using a citalopram-derived affinity resin

Journal Article · Tue Apr 03 00:00:00 EDT 1990 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00465a029· OSTI ID:6073899

Biessen, E A L; Horn, A S; Robillard, G T ^[1]

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1. Univ. of Groningen (Netherlands)

This paper describes a procedure for the synthesis and application of a citalopram-derived affinity resin in purifying the 5HT-reuptake system from human blood platelets. A two-step scheme has been developed for partial purification, based on wheat germ agglutinin-lectin (WGA) affinity and citalopram affinity chromatographies. Upon solubilization of the carrier with 1% digitonin, a 50-70-fold increase in specific ({sup 3}H) imipramine binding activity with a 70% recovery could be accomplished through WGA-lectin chromatography. The WGA pool was then subjected to affinity chromatography on citalopram-agarose. At least 90% of the binding capacity adsorbed to the column. Specific elution using 10 {mu}M citalopram resulted in a 22% recovery of binding activity. A 10,000-fold overall purification was obtained by using this two-step procedure. Analysis of the fractions on SDS-PAGE after {sup 125}I labeling revealed specific elution of 78- and 55-kDa proteins concomitant with the appearance of ({sup 3}H) imipramine binding activity. The pharmacological profile of the partially purified reuptake system correlated well with that derived from the crude membrane-bound reuptake system, suggesting a copurification of the 5HT binding activity and ({sup 3}H)imipramine binding activity.

• OSTI ID: 6073899
• Journal Information: Biochemistry; (USA), Vol. 29:13; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
IMIPRAMINE
UPTAKE
MEMBRANE PROTEINS
PURIFICATION
TRITIUM COMPOUNDS
BLOOD PLATELETS
ELECTROPHORESIS
HYDROXYTRYPTOPHAN
IODINE 125
MAN
SODIUM IODIDES
ALKALI METAL COMPOUNDS
AMINES
AMINO ACIDS
ANIMALS
ANTIDEPRESSANTS
BETA DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CENTRAL NERVOUS SYSTEM AGENTS
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
HALIDES
HALOGEN COMPOUNDS
HETEROCYCLIC COMPOUNDS
HYDROGEN COMPOUNDS
HYDROXY ACIDS
INORGANIC PHOSPHORS
INTERMEDIATE MASS NUCLEI
IODIDES
IODINE COMPOUNDS
IODINE ISOTOPES
ISOTOPES
MAMMALS
MATERIALS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORS
PRIMATES
PROTEINS
PSYCHOTROPIC DRUGS
RADIOISOTOPES
RADIOPROTECTIVE SUBSTANCES
SODIUM COMPOUNDS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques