Adenosine deaminase in cell transformation. Biophysical manifestation of membrane dynamics.
Cell transformation is associated with a dramatic collapse of a graphic fingerprint characteristic of normal cells, as measured by phase fluorimetry. This is demonstrated on adenosine deaminase (ADA, EC 3.5.4.4), an established malignancy marker. ADA activity is known to decrease markedly in chick embryo fibroblasts (CEF) transformed by Rous sarcoma virus. The high affinity between the catalytic small subunit ADA (SS-ADA) and its membranal complexing protein (ADCP) (which abounds on the plasma membrane of CEF) allowed the hybridization of fluorescent labeled SS-ADA with native ADCP on CEF. Multifrequency differential phase fluorimetry responded remarkably to the state of this hybrid membrane protein. The transformation process is shown to have led to increased membrane fluidity and rotational mobility of ADCP as well as to its reduced availability to SS-ADA binding. The hypothesis of protein vertical sinking into the lipid core of the membrane is now given support by our spectroscopic data. Additional models are considered. A regulatory role is thus suggested for the complexing protein, which may also account for (a) reduced ADA activity in transformed cells and (b) detachment, exclusive to normal cells, upon addition of SS-ADA in excess.
Porat
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N
Department of Chemistry, Ben-Gurion University of the Negev, Beer Sheva, Israel.
Gill
D
D
Parola
A H
AH
D000243:Adenosine Deaminase / Q000378:metabolism*; D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology*; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D002642:Chick Embryo; D006023:Glycoproteins / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008856:Microscopy, Fluorescence; D009700:Nucleoside Deaminases / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
D006023:Glycoproteins; D007527:Isoenzymes; D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase