TY - JOUR
T1 - Affinity chromatography of heavy meromyosin subfragment-1 reacted with thiol reagents
AU - Lamed, Raphael
AU - Oplatka, Avraham
AU - Reisler, Emil
N1 - Funding Information:
The work of A.O. was supported by a grant from the Muscular Dystrophy Associations of America.
PY - 1976/4/14
Y1 - 1976/4/14
N2 - Separation of heavy meromyosin subfragment-1 treated with N-ethyl maleimide (Ma1NEt) into native -SH1- and - (SH1,SH2)-blocked protein populations could be achieved by affinity chromatography on agarose-ATP columns in the presence of Mg2+ or Ca2+. Covalent bridging of the two -SH groups by p-phenylenedimaleimide gave a product which has the same affinity of binding to ATP columns as the doubly blocked Ma1NEt preparation. Treatment with p-phenylenedimaleimide abolished binding to immobilized F-actin columns, whereas modifications by Ma1NEt did not affect adsorption by this chromatographic medium. Affinity chromatography on immobilized nucleotide and actin columns is suggested as an analytical tool in the study of the involvement of thiol groups in the myosin active site and its conformation.
AB - Separation of heavy meromyosin subfragment-1 treated with N-ethyl maleimide (Ma1NEt) into native -SH1- and - (SH1,SH2)-blocked protein populations could be achieved by affinity chromatography on agarose-ATP columns in the presence of Mg2+ or Ca2+. Covalent bridging of the two -SH groups by p-phenylenedimaleimide gave a product which has the same affinity of binding to ATP columns as the doubly blocked Ma1NEt preparation. Treatment with p-phenylenedimaleimide abolished binding to immobilized F-actin columns, whereas modifications by Ma1NEt did not affect adsorption by this chromatographic medium. Affinity chromatography on immobilized nucleotide and actin columns is suggested as an analytical tool in the study of the involvement of thiol groups in the myosin active site and its conformation.
UR - http://www.scopus.com/inward/record.url?scp=0017250764&partnerID=8YFLogxK
U2 - 10.1016/0005-2795(76)90212-9
DO - 10.1016/0005-2795(76)90212-9
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C2 - 131579
AN - SCOPUS:0017250764
SN - 0005-2795
VL - 427
SP - 688
EP - 695
JO - BBA - Protein Structure
JF - BBA - Protein Structure
IS - 2
ER -