Hydrolytically induced allosteric change in the heavy chain of intact myosin involving nonessential thiol groups

• Published in: Biochemistry (Easton) 1978-01, Vol.17 (2), p.246-253
• Main Authors: Schaub, Marcus C, Watterson, John G, Walser, Juerg T, Waser, Peter G
• Format: Article
• Language: English
• Subjects: Adenine Nucleotides - metabolism; Allosteric Regulation; Amino Acid Sequence; Binding Sites; Chemical Phenomena; Chemistry; Ethylmaleimide; Hydrolysis; Molecular Weight; Myosin Subfragments - metabolism; Myosins - metabolism; Peptide Fragments - metabolism; Protein Conformation; Space life sciences; Sulfhydryl Compounds - analysis
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00595a009

The two globular head portions, each bearing an active site, contain an uncleaved heavy chain when isolated by chymotrypsin from intact myosin. By specific labeling with radioactive N-ethylmaleimide the essential thiol 1 and thiol 2 groups were found to reside in this heavy chain. In intact myosin nonessential thiol 3 groups become the most reactive during ATP hydrolysis above 15 degrees C. These thiol 3 groups are located in a portion of the myosin heavy chain which appears as a fragment with an apparent molecular weight of 11 000 during proteolysis. The facts that this fragment is produced in an almost 1: 1 molar ratio with the head heavy chain and that it bears unblocked N-terminal amino groups whereas the heavy chain does not and is not contained in the rod portion of the myosin molecule indicate that it may orginate from the heavy chains in the neck region where the heads are joined to the rod. Since this fragment is removed by ion-exchange chromatography, it is not part of the functioning head and hence not involved in the active site. As its nonessential thiol 3 groups are rendered the most reactive of all thiol groups in the enzyme-product complex M**ADP.Pi, the hydrolytic step induces an allosteric conformational change in the neck region of intact myosin.