The magnesium-ATP-binding site on chicken gizzard myosin light chain kinase remains open and functionally competent during the calmodulin-dependent activation-inactivation cycle of the enzyme

Biochemistry ◽  
1992 ◽  
Vol 31 (23) ◽  
pp. 5394-5399 ◽  
Author(s):  
Peter J. Kennelly ◽  
Jie Leng ◽  
Petra Marchand
Keyword(s):  
Binding Site ◽  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Atp Binding ◽  
Atp Binding Site ◽  
Chicken Gizzard

scholarly journals Affinity labelling of smooth-muscle myosin light-chain kinase with 5′-[p-(fluorosulphonyl)benzoyl]adenosine

1993 ◽  
Vol 296 (1) ◽  
pp. 53-58 ◽  
Author(s):  
H Komatsu ◽  
M Ikebe
Keyword(s):  
Smooth Muscle ◽  
Binding Site ◽  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Smooth Muscle Myosin ◽  
Atp Binding ◽  
Atp Binding Site ◽  
Calmodulin Binding ◽  
Muscle Myosin

5′-(p-(Fluorosulphonyl)[14C]benzoyl)adenosine (FSBA) was synthesized and used as a probe to study the ATP-binding site of smooth-muscle myosin light-chain kinase (MLCK). FSBA modified both free MLCK and calmodulin/MLCK complex, resulting in inactivation of the kinase activity. Nearly complete protection of the calmodulin/MLCK complex against FSBA modification was obtained by addition of excess ATP whereas MLCK activity alone was lost in a dose-dependent manner even in the presence of excess ATP. These results suggest that FSBA modified ATP-binding sites and ATP-independent sites, and the latter sites are protected by calmodulin binding. The results also suggest that the ATP-binding site is accessible to the nucleotide substrate regardless of calmodulin binding. The FSBA-labelled MLCK was completely proteolysed by alpha-chymotrypsin, and the 14C-labelled peptides were isolated and sequenced. The sequence of the labelled peptide was Ala-Gly-X-Phe, where X is the labelled residue. The sequence was compared with the known MLCK sequence, and the labelled residue was identified as lysine-548, which is located downstream of the GXGXXG motif conserved among ATP-utilizing enzymes.

scholarly journals Isolation of the native form of chicken gizzard myosin light-chain kinase

1984 ◽  
Vol 218 (3) ◽  
pp. 863-870 ◽  
Author(s):  
P K Ngai ◽  
C A Carruthers ◽  
M P Walsh
Keyword(s):  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Binding Protein ◽  
Cross Reactivity ◽  
Native Form ◽  
Chicken Gizzard ◽  
Calmodulin Binding ◽  
Chymotryptic Peptide ◽  
Dependent Protein

A simple and rapid procedure for the purification of the native form of chicken gizzard myosin light-chain kinase (Mr 136000) is described which eliminates problems of proteolysis previously encountered. During this procedure, a calmodulin-binding protein of Mr 141000, which previously co-purified with the myosin light-chain kinase, is removed and shown to be a distinct protein on the basis of lack of kinase activity, different chymotryptic peptide maps, lack of cross-reactivity with a monoclonal antibody to turkey gizzard myosin light-chain kinase, and lack of phosphorylation by the purified catalytic subunit of cyclic AMP-dependent protein kinase. This Mr-141000 calmodulin-binding protein is identified as caldesmon on the basis of Ca2+-dependent interaction with calmodulin, subunit Mr, Ca2+-independent interaction with skeletal-muscle F-actin, Ca2+-dependent competition between calmodulin and F-actin for caldesmon, and tissue content.

Modulator protein as a component of the myosin light chain kinase from chicken gizzard

Biochemistry ◽  
1978 ◽  
Vol 17 (2) ◽  
pp. 253-258 ◽  
Author(s):  
Renata Dabrowska ◽  
James M. F. Sherry ◽  
Debra K. Aromatorio ◽  
David J. Hartshorne
Keyword(s):  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Chicken Gizzard

scholarly journals Regulatory and structural motifs of chicken gizzard myosin light chain kinase.

1990 ◽  
Vol 87 (6) ◽  
pp. 2284-2288 ◽  
Author(s):  
N. J. Olson ◽  
R. B. Pearson ◽  
D. S. Needleman ◽  
M. Y. Hurwitz ◽  
B. E. Kemp ◽  
...  
Keyword(s):  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Structural Motifs ◽  
Chicken Gizzard

Domain organization of chicken gizzard myosin light chain kinase deduced from a cloned cDNA

Biochemistry ◽  
1986 ◽  
Vol 25 (26) ◽  
pp. 8372-8381 ◽  
Author(s):  
Vince Guerriero ◽  
Mario A. Russo ◽  
Norma J. Olson ◽  
John A. Putkey ◽  
Anthony R. Means
Keyword(s):  
Light Chain ◽  
Myosin Light Chain Kinase ◽  
Myosin Light Chain ◽  
Domain Organization ◽  
Chicken Gizzard ◽  
Cloned Cdna
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