Covalent modification of the amine transporter with N,N prime -dicyclohexylcarbodiimide

Journal Article · Tue Jul 02 04:00:00 EDT 1991 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00240a020· OSTI ID:5032361

Suchi, R; Stern-Bach, Y; Gabay, T; Schuldiner, S ^[1]

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1. Hebrew Univ. of Jerusalem (Israel)

N,N{prime}-Dicyclohexylcarbodiimide (DCC) has been previously shown to inhibit the amine transporter from chromaffin granules. A study of the mechanism of inhibition is present together with the demonstration of covalent modification of the protein. DCC inhibits binding of R1 (reserpine) and R2 (tetrabenazine) types of ligands to the transporter as well as transport. Lignads of the R2 type, but not those of the R1 type, protect against inhibition of all the reactions by DCC, i.e., accumulation of serotonin, binding if reserpine (R1 ligand), and binding of ketanserine (R2 ligand). The ability of a given R2 ligand to protect the transporter correlates well with its binding constant. Water-soluble carbodiimides, such as 1-ethyl-3-(3-(diethylamino)propyl)carbodiimide (EDC), do not have any effect on the catalytic activity of the transporter. A flourescent hydrophobic analogue of DCC, N-cyclohexyl-N{prime}-(4-(dimethylamino)-{alpha}-naphthyl)carbodiimide (NCD-4), inhibits at about the same concentration range as DCC. ({sup 14}C)DCC labels several polypeptides in the chromaffin granule membranes. Labeling of a polypeptide with an apparent M{sub r} of 80K is inhibited in the presence of R2 ligands. The labeled polypeptide copurifies with the recently identified and isolated transporter.

• OSTI ID: 5032361
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 30:26; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALOIDS
AMINES
ANTIHYPERTENSIVE AGENTS
AROMATICS
AUTONOMIC NERVOUS SYSTEM AGENTS
AZAARENES
AZOLES
CARBON 14 COMPOUNDS
CARBON COMPOUNDS
CARDIOVASCULAR AGENTS
DRUGS
HETEROCYCLIC COMPOUNDS
HYDROGEN COMPOUNDS
INDOLES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIGANDS
MEMBRANE PROTEINS
MEMBRANE TRANSPORT
MOLECULAR STRUCTURE
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PH VALUE
PROTEINS
PURIFICATION
PYRROLES
RADIORECEPTOR ASSAY
RESERPINE
SYMPATHOLYTICS
TRACER TECHNIQUES
TRITIUM COMPOUNDS