The interaction of 5'-nucleotidase purified from chicken gizzard and actin, and the reversible loss of the inhibitory capacity of actin on deoxyribonuclease I.
Evidence is presented for a direct interaction of the intrinsic membrane protein 5'-nucleotidase (5'-ribonucleotide phosphohydrolase, EC 3.1.3.5) purified from avian smooth muscle (chicken gizzard) and the cytoskeletal component actin. Two different modes of interaction can be discerned: firstly, an immediate inhibitory effect of preferentially filamentous actin (F-actin) on the enzymic (i.e., AMPase) activity of 5'-nucleotidase and a direct binding of this enzyme to immobilized F-actin. Since these effects are suppressed by the addition of myosin subfragment 1, binding of 5'-nucleotidase appears to occur along the F-actin filament axis. Secondly, a time- and 5'-nucleotidase concentration-dependent transformation of also preferentially F-actin into a form unable to inhibit the enzymic activity of deoxyribonuclease I (DNAase I). This desensitization of actin versus DNAase I is not due to a denaturation process and was found to be reversible after addition of ATP. Furthermore, it does not seem to effect the ability of actin to bind to DNAase I. The transformation is accompanied by the hydrolysis of actin-bound nucleotide into adenosine, which remains bound to actin. Therefore, the desensitization of actin versus DNAase I appears to be due to a nucleotide-dependent conformational change of actin. An unidentified contamination of the 5'-nucleotidase preparations to a varying degree with ADPase and ATPase activities appears to be responsible for the desensitization process, although a synergistic role of these activities and 5'-nucleotidase cannot be excluded.
Dieckhoff
J
J
Mannherz
H G
HG
D015720:5'-Nucleotidase; D000199:Actins / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D002645:Chickens; D003850:Deoxyribonuclease I / Q000037:antagonists & inhibitors*; D005895:Gizzard, Avian / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D009708:Nucleotidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011108:Polymers / Q000378:metabolism
D000199:Actins; D011108:Polymers; D003850:Deoxyribonuclease I; D009708:Nucleotidases; D015720:5'-Nucleotidase