Functional reconstitution of the olfactory membrane: incorporation of the olfactory adenylate cyclase in liposomes.

Abstract:

:Chemosensory cilia isolated from the olfactory epithelium of Rana catesbeiana were solubilized with Lubrol PX in the presence of supplementary lipid, forskolin, and sodium fluoride. Subsequent removal of the detergent by adsorption onto Biobeads SM2 results in the formation of proteoliposomes that display forskolin- and GTP gamma S-sensitive adenylate cyclase activity. Sucrose gradient centrifugation of liposomes formed in the presence of fluorescently labeled phosphatidylcholine demonstrates association between the olfactory adenylate cyclase and the exogenously added lipid. Forskolin stimulates the enzyme in reconstituted membranes with the same potency as in native membranes (EC50 = 1-2 microM). However, GTP gamma S is 350-fold more potent in native membranes (EC50 = 4.0 +/- 0.5 nM) than in reconstituted membranes (EC50 = 1.4 +/- 0.3 microM). These studies represent a first step toward the functional reconstitution and molecular dissection of the olfactory membrane.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Anholt RR

doi

10.1021/bi00417a040

subject

Has Abstract

pub_date

1988-08-23 00:00:00

pages

6464-8

issue

17

eissn

0006-2960

issn

1520-4995

journal_volume

27

pub_type

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