4-Azido-2-nitrophenyl phosphate, a new photoaffinity derivative of inorganic phosphate. Study of its interaction with the inorganic phosphate binding site of beef heart mitochondrial adenosine triphosphatase

Biochemistry ◽  
1980 ◽  
Vol 19 (20) ◽  
pp. 4620-4626 ◽  
Author(s):  
Guy Lauquin ◽  
Richard Pougeois ◽  
Pierre V. Vignais
Keyword(s):  
Binding Site ◽  
Inorganic Phosphate ◽  
Adenosine Triphosphatase ◽  
Beef Heart ◽  
Phosphate Binding ◽  
Nitrophenyl Phosphate

Further investigations on the inorganic phosphate binding site of beef heart mitochondrial F1-ATPase

Biochemistry ◽  
1985 ◽  
Vol 24 (4) ◽  
pp. 1020-1024 ◽  
Author(s):  
Richard Pougeois ◽  
Guy J. M. Lauquin
Keyword(s):  
Binding Site ◽  
Inorganic Phosphate ◽  
Beef Heart ◽  
Phosphate Binding ◽  
F1 Atpase

Phosphate Binding to Isolated Chloroplast Coupling Factor (CF1)

1988 ◽  
Vol 43 (3-4) ◽  
pp. 213-218 ◽  
Author(s):  
Bernhard Huchzermeyer
Keyword(s):  
Atpase Activity ◽  
Binding Site ◽  
Inorganic Phosphate ◽  
Coupling Factor ◽  
Binding Domain ◽  
Atp Binding ◽  
Phosphate Binding ◽  
Chloroplast Coupling Factor ◽  
Single Binding Site ◽  
Site Binding

A single binding site for phosphate was found on isolated chloroplast coupling factor in the absence of nucleotides. In our experiments the phosphate binding site showed a Kd of 170 μᴍ. We did not observe any differences whether the ATPase activity of CF] had been activated or not. If the enzyme was incubated with [γ-32P]ATP the amount of 32P bound per CF1 depended on the pretreatment of the enzyme: In the presence of ADP no ATP or phosphate was bound to CF,. After activation of ATPase activity one mol of ATP per mol CF, was rapidly bound and hydrolyzed while there was a slowly occurring binding of another phosphate without concomitant nucleotide binding. We conclude that there are two different types of phosphate binding observed in our experiments: 1) Inorganic phosphate can be bound by one catalytic site per mol of CF1 2) The γ-phosphate of ATP is able to bind to an ATP binding domain of the enzyme if this domain can exchange substrates with the incubation medium. This ATP binding domain appears to differ from the site binding inorganic phosphate, because at least a portion of the coupling factor contains more than one labelled phosphate during our ATPase tests.

scholarly journals Allosteric interactions of glycogen phosphorylase b. A crystallographic study of glucose 6-phosphate and inorganic phosphate binding to di-imidate-cross-linked phosphorylase b

1984 ◽  
Vol 218 (1) ◽  
pp. 45-60 ◽  
Author(s):  
A Lorek ◽  
K S Wilson ◽  
M S P Sansom ◽  
D I Stuart ◽  
E A Stura ◽  
...  
Keyword(s):  
Binding Site ◽  
Conformational Changes ◽  
Inorganic Phosphate ◽  
Glycogen Phosphorylase ◽  
Phosphate Binding ◽  
Binding Studies ◽  
Structural Explanation ◽  
Alpha Helices ◽  
Allosteric Effector ◽  
Glycogen Phosphorylase B

The binding to glycogen phosphorylase b of glucose 6-phosphate and inorganic phosphate (respectively allosteric inhibitor and substrate/activator of the enzyme) were studied in the crystal at 0.3 nm (3A) resolution. Glucose 6-phosphate binds in the alpha-configuration at a site that is close to the AMP allosteric effector site at the subunit-subunit interface and promotes several conformational changes. The phosphate-binding site of the enzyme for glucose 6-phosphate involves contacts to two cationic residues, Arg-309 and Lys-247. This site is also occupied in the inorganic-phosphate-binding studies and is therefore identified as a high-affinity phosphate-binding site. It is distinct from the weaker phosphate-binding site of the enzyme for AMP, which is 0.27 nm (2.7A) away. The glucose moiety of glucose 6-phosphate and the adenosine moiety of AMP do not overlap. The results provide a structural explanation for the kinetic observations that glucose 6-phosphate inhibition of AMP activation of phosphorylase b is partially competitive and highly co-operative. The results suggest that the transmission of allosteric conformational changes involves an increase in affinity at phosphate-binding sites and relative movements of alpha-helices. In order to study glucose 6-phosphate and phosphate binding it was necessary to cross-link the crystals. The use of dimethyl malondi-imidate as a new cross-linking reagent in protein crystallography is discussed.

scholarly journals AlF4- reversibly inhibits ‘P’-type cation-transport ATPases, possibly by interacting with the phosphate-binding site of the ATPase

1988 ◽  
Vol 253 (3) ◽  
pp. 827-833 ◽  
Author(s):  
L Missiaen ◽  
F Wuytack ◽  
H De Smedt ◽  
M Vrolix ◽  
R Casteels
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Binding Site ◽  
G Proteins ◽  
Time Course ◽  
Cation Transport ◽  
Phosphate Binding ◽  
Intact Cells ◽  
Nitrophenyl Phosphate ◽  
Transport Atpases ◽  
P Type

The only known cellular action of AlF4- is to stimulate the G-proteins. The aim of the present work is to demonstrate that AlF4- also inhibits ‘P’-type cation-transport ATPases. NaF plus AlCl3 completely and reversibly inhibits the activity of the purified (Na+ + K+)-ATPase (Na+- and K+-activated ATPase) and of the purified plasmalemmal (Ca2+ + Mg2+)-ATPase (Ca2+-stimulated and Mg2+-dependent ATPase). It partially inhibits the activity of the sarcoplasmic-reticulum (Ca2+ + Mg2+)-ATPase, whereas it does not affect the mitochondrial H+-transporting ATPase. The inhibitory substances are neither F- nor Al3+ but rather fluoroaluminate complexes. Because AlF4- still inhibits the ATPase in the presence of guanosine 5′-[beta-thio]diphosphate, and because guanosine 5′-[beta gamma-imido]triphosphate does not inhibit the ATPase, it is unlikely that the inhibition could be due to the activation of an unknown G-protein. The time course of inhibition and the concentrations of NaF and AlCl3 required for this inhibition differ for the different ATPases. AlF4- inhibits the (Na+ + K+)-ATPase and the plasmalemmal (Ca2+ + Mg2+)-ATPase noncompetitively with respect to ATP and to their respective cationic substrates, Na+ and Ca2+. AlF4- probably binds to the phosphate-binding site of the ATPase, as the Ki for inhibition of the (Na+ + K+)-ATPase and of the plasmalemmal (Ca2+ + Mg2+)-ATPase is shifted in the presence of respectively 5 and 50 mM-Pi to higher concentrations of NaF. Moreover, AlF4- inhibits the K+-activated p-nitrophenylphosphatase of the (Na+ + K+)-ATPase competitively with respect to p-nitrophenyl phosphate. This AlF4- –induced inhibition of ‘P’-type cation-transport ATPases warns us against explaining all the effects of AlF4- on intact cells by an activation of G-proteins.

Crystal Structure of Glycosomal Glyceraldehyde-3-phosphate Dehydrogenase from Leishmania mexicana: Implications for Structure-Based Drug Design and a New Position for the Inorganic Phosphate Binding Site

Biochemistry ◽  
1995 ◽  
Vol 34 (46) ◽  
pp. 14975-14986 ◽  
Author(s):  
Hidong Kim ◽  
Ingeborg K. Feil ◽  
Christophe L. M. J. Verlinde ◽  
Philip H. Petra ◽  
Wim G. J. Hol
Keyword(s):  
Crystal Structure ◽  
Drug Design ◽  
Binding Site ◽  
Inorganic Phosphate ◽  
Leishmania Mexicana ◽  
Phosphate Binding ◽  
Structure Based Drug Design

Characterization of an inorganic phosphate binding site on the isolated, reconstitutively active .beta. subunit of F0.cntdot.F1 ATP synthase

Biochemistry ◽  
1985 ◽  
Vol 24 (10) ◽  
pp. 2482-2487 ◽  
Author(s):  
Daniel Khananshvili ◽  
Zippora Gromet-Elhanan
Keyword(s):  
Binding Site ◽  
Atp Synthase ◽  
Inorganic Phosphate ◽  
Beta Subunit ◽  
Phosphate Binding

scholarly journals Periodate-oxidized AMP as a substrate, an inhibitor and an affinity label of human placental alkaline phosphatase

1981 ◽  
Vol 199 (2) ◽  
pp. 281-287 ◽  
Author(s):  
G G Chang ◽  
S C Wang ◽  
F Pan
Keyword(s):  
Alkaline Phosphatase ◽  
Inorganic Phosphate ◽  
Competitive Inhibitor ◽  
Binary Complex ◽  
Placental Alkaline Phosphatase ◽  
Phosphate Binding ◽  
Complete Inactivation ◽  
Nitrophenyl Phosphate ◽  
Saturation Kinetics ◽  
Human Placental Alkaline Phosphatase

Human placental alkaline phosphatase (EC 3.1.3.1) was inactivated by periodate-oxidized AMP. The inactivation showed saturation kinetics and could be partially prevented by the substrate AMP or the product inhibitor inorganic phosphate. Oxidized AMP was itself a substrate for this enzyme, with an apparent Km of 0.67 mM. The hydrolytic products of oxidized AMP were identified as oxidized adenosine hemiacetals. Oxidized AMP was also found to be a non-competitive inhibitor with respect to p-nitrophenyl phosphate, with identical Kis and Kii values of 0.15 mM. Our results indicate that oxidized AMP could combine with the enzyme to form a binary complex, followed by reaction with the proximal lysyl amino group to yield a Schiff base. The latter was reduced with NaBH4 and identified by t.l.c. The incorporation of only 1.5 molecules of oxidized [14C]AMP per enzyme subunit resulted in a complete inactivation of the enzyme. The modified enzyme showed higher apparent Km for the substrates and higher Ki for inorganic phosphate, but lower [32P]phosphate incorporation, than the native enzyme. These results support the conclusion that a lysine residue is involved in the phosphate-binding site of human placental alkaline phosphatase.

scholarly journals Reversible binding of Pi by beef heart mitochondrial adenosine triphosphatase.

1977 ◽  
Vol 252 (9) ◽  
pp. 2891-2899 ◽  
Author(s):  
H S Penefsky
Keyword(s):  
Adenosine Triphosphatase ◽  
Beef Heart ◽  
Reversible Binding

scholarly journals The Subunit Structure of Beef Heart Mitochondrial Adenosine Triphosphatase

1972 ◽  
Vol 247 (20) ◽  
pp. 6617-6623 ◽  
Author(s):  
Aileen F. Knowles ◽  
Harvey S. Penefsky
Keyword(s):  
Adenosine Triphosphatase ◽  
Beef Heart ◽  
Subunit Structure
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