The Use of MRNA Translation in Vitro and in Ovo Followed by Crossed Immunoelectrophoretic Autoradiography to Study the Biosynthesis of Human Cholinesterases

Overview

Journal Cell Mol Neurobiol

Publisher Springer

Specialties Cell Biology
Molecular Biology
Neurology

Date 1986 Sep 1

PMID 3802131

Citations 2

Authors

H Soreq

K M Dziegielewska

D Zevin-Sonkin

H Zakut

Affiliations

Soon will be listed here.

Abstract

The synthesis of various cholinesterases in different fetal human tissues was studied using in vitro and in ovo translation of poly(A)+ RNA, followed by crossed immunoelectrophoretic autoradiography. When unfractionated poly(A)+ mRNA from fetal brain, muscle, or liver was translated in vitro, in the reticulocyte lysate cell-free system, polypeptides were synthesized which reacted with antibodies against either "true" acetylcholinesterase (acetylcholine hydrolase; EC 3.1.1.7) or "pseudo", butyrylcholinesterase (acylcholine acylhydrolase; EC 3.1.1.8). The two nascent cholinesterases could be separated by crossed immunoelectrophoresis followed by autoradiography, suggesting that acetylcholinesterase and butyrylcholinesterase are produced in all three tissues from nascent polypeptides containing different immunological domains. To examine whether the biosynthesis of cholinesterases includes posttranslational processing events, Xenopus oocytes were microinjected with mRNA from these tissues. Immunoelectrophoretic analysis of oocyte intracellular homogenates and incubation medium revealed various precipitation arcs, reflecting the synthesis and posttranslational processing of multiple forms of tissue-specific exported and intracellular acetylcholinesterase and butyrylcholinesterase. These findings demonstrate that polymorphic cholinesterases are produced from nascent polypeptide products which undergo further posttranslational processing events in a tissue-specific manner before they become mature compartmentalized cholinesterases.

Citing Articles

[The development of pharmacogenetics--a retrospective on the 75th birthday of Hans Herken].

KALOW W Klin Wochenschr. 1988; 66(6):229-35.

PMID: 3285092 DOI: 10.1007/BF01748161.

Molecular biological search for human genes encoding cholinesterases.

Soreq H, Gnatt A Mol Neurobiol. 1987; 1(1-2):47-80.

PMID: 3077058 DOI: 10.1007/BF02935264.

Tissue-specific processing and polarized compartmentalization of clone-produced cholinesterase in microinjected Xenopus oocytes.

Dreyfus P, Seidman S, Pincon-Raymond M, Murawsky M, Rieger F, Schejter E Cell Mol Neurobiol. 1989; 9(3):323-41.

PMID: 2692828 PMC: 11567229. DOI: 10.1007/BF00711413.

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