Preparation and characterization of mitochondrial myosins of rat and human liver.
This paper confirmed the reality of the mitochondrial myosin (mt-myosin in human and rat liver. Simultaneously, cytoplasmic myosin (cp-myosin) was prepared from the large particle-free supernatant. The yield of purified mt- and cp-myosin from 1 kg fresh liver was altogether 5-600 mg (= 1-1.2 mumol). Half of the myosin originated from the mitochondrial fraction (composed of about 60 g of mitochondrial protein), while the remaining portion (cp-myosin) was derived from a translucent, but voluminous supernatant (containing about two-third of liver proteins). Comparing the molecular mass of mt- and cp-myosin to the skeletal muscle myosin (about 480 kDa)--on the basis of gel filtration profiles--they proved to have similar profiles. The characteristic properties of both preparations were similar to other myosins developing filamentous aggregations and showing ATP-induced superprecipitations, but they had lower ATPase activities thus being more similar to smooth muscle and cell myosins than to skeletal muscle myosin. The mitochondria and both myosins contained abundant covalently bound P and their endogeneous Si content was low, 2-3 mumol/g in fresh mitochondria and 5-7 mol Si per mol in the mt-myosin. The Si content was resolved into 2-3, while P into 5-6 fractions as revealed by ion exchange chromatographic technique. The mt-myosin could be saturated to a higher P level by autophosphorylation than the cytoplasmic myosin. The interaction of actin with myosin induced a release of significant amounts of P, depending on the ATP concentration. The Cu2+ treatment of mt-myosin caused also P release, and a limited amount of Cu remained bound in the preparations.
2nd Institute of Biochemistry, Semmelweis University Medical School, Budapest, Hungary.
S
S
Fazekas
J
J
Fehér
L
L
Kondics
V
V
Székessy-Hermann
D000199:Actins / Q000494:pharmacology; D000818:Animals; D006801:Humans; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D008930:Mitochondria, Liver / Q000201:enzymology*; D009218:Myosins / Q000032:analysis* / Q000378:metabolism; D010710:Phosphates / Q000032:analysis; D010766:Phosphorylation; D051381:Rats
D000199:Actins; D007527:Isoenzymes; D010710:Phosphates; D009218:Myosins