Characterization of stable beryllium fluoride, aluminum fluoride, and vanadate containing myosin subfragment 1-nucleotide complexes.
Beryllium and aluminum fluorides are good phosphate analogues. These compounds, like orthovanadate, form stable complexes with myosin subfragment 1 (S1) in the presence of MgADP. The formation of the stable S1-nucleotide complexes is characterized by the loss of ATPase activity. For the complete loss of ATPase activity there was necessary a higher concentration of aluminum than of beryllium or vanadate. In the presence of MgATP the onset of the inhibition is delayed, which indicates that stable complexes cannot form when a specific site is occupied by the gamma-phosphate of ATP or by P(i) derived from the gamma-phosphate. The half-lives of the S1-MgADP-(BeF3-), S1-MgADP-(AlF4-), and S1-MgADP-Vi complexes at 0 degrees C are 7, 2, and 4 days, respectively. In the presence of actin the rate of decomposition of all of the complexes is significantly enhanced; however, the order of decomposition is reversed, the fastest rate being observed with beryllium and the slowest with aluminum. The formation of the S1-MgADP-(BeF3-) and S1-MgADP-(AlF4-) complexes is accompanied by an increase in tryptophan fluorescence similar to that observed upon addition of MgATP to S1. The fluorescence increase develops rather slowly, by suggesting that the rate-limiting step in the formation of the stable complex is an isomerization. The rate of the fluorescence change accompanying the formation of the Be complex is faster than that for the Al complex. Addition of vanadate to S1 causes a static quenching of the tryptophan fluorescence.(ABSTRACT TRUNCATED AT 250 WORDS)
Werber
M M
MM
Department of Oral Biology, Hebrew University-Hadassah School of Dental Medicine, Jerusalem, Israel.
Peyser
Y M
YM
Muhlrad
A
A
D000199:Actins / Q000737:chemistry; D000244:Adenosine Diphosphate / Q000737:chemistry; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000535:Aluminum / Q000737:chemistry*; D017607:Aluminum Compounds; D000818:Animals; D001608:Beryllium / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005459:Fluorides / Q000737:chemistry*; D007700:Kinetics; D015879:Myosin Subfragments / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism*; D011817:Rabbits; D013696:Temperature; D014638:Vanadates / Q000737:chemistry*
D000199:Actins; D017607:Aluminum Compounds; D015879:Myosin Subfragments; D009711:Nucleotides; D014638:Vanadates; C035982:beryllium fluoride; D000244:Adenosine Diphosphate; D000535:Aluminum; D000251:Adenosine Triphosphatases; D001608:Beryllium; D005459:Fluorides; C032311:aluminum fluoride