Isolation of Sperm Whale Myoglobin by Low Temperature Fractionation with Ethanol and Metallic Ions

Hardman, Karl D.; Eylar, E.H.; Ray, Dilip Kumar; Banaszak, Leonard J.; Gurd, Frank R. N.

doi:10.1016/s0021-9258(18)96936-x

Cited by 122 publications

(10 citation statements)

References 33 publications

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“…However, the acid range of the titration curves (Figure 4) is more informative since components A|, A2Ci, and A2C2 align with the natural sperm whale myoglobin component IV, A3 and A4 coincide with the natural minor component III (not shown), and A5 aligns with the natural minor component II. The minor components III and II are 1 and 2 charges, respectively, more negative than the major IV (Hardman et al, 1966;Nakhleh, 1971). These results suggest that the last three acetimidyl components, unlike the first three, a Protein hydrolysis was performed for 24, 48, and 72 h. Serine, threonine, and acetimidolysine content were determined by extrapolation to zero time while isoleucine and valine contents were determined by extrapolation to 100 h. * The yield reported under sequencer cycle is that fraction of N-terminal remaining unreacted.…”

Section: Resultsmentioning

confidence: 99%

Characterization of the reaction of methyl acetimidate with sperm whale myoglobin

DiMarchi¹,

Garner²,

Wang³

et al. 1978

Biochemistry

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The effects of pH, acetimidate concentration, temperature, and reaction time of methyl acetimidate with sperm whale myoglobulin have been assessed. Reaction at pH 9.8 and 15 degrees C for 30 min with a sixfold excess of methyl acetimidate relative to each amino group yielded six acetimidomyoglobin derivatives which were separated and purified. Reaction with tetrahydrophthalic anhydride revealed the number of amino groups that remained unreacted in each separated component and made possible further subractionation. Modification at the NH2 terminus was quantitated by automated stepwise Edman degradation. The acetimidyl and tetrahydrophthalyl groups, were readily removable. The potentiometric titration of three of the completely deprotected components showed identity with the parent untreated sperm whale myoglobin. The first of two major products was acetimidated at all 19 epsilon-amino groups but not at the NH2 terminus. The second major product bore a blocked NH2 terminus but retained one unmodified epsilon-amino group, identified after modification by trinitrobenzenesulfonate as lysine residue 77. Of the minor components, one was identified as completely acetimidated at all 20 amino groups. The other three minor components appeared to contain irreversible by-products.

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Section: Resultsmentioning

confidence: 99%

Characterization of the reaction of methyl acetimidate with sperm whale myoglobin

DiMarchi¹,

Garner²,

Wang³

et al. 1978

Biochemistry

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“…The primary structure of all synthetic intermediates was ascertained through amino acid analysis, NH2-terminal sequence quantitation, and electrophoresis. Once 0 Wavelength absorbance ratios as determined by Nakhleh (1971) and Hardman et al (1966). and homogeneity of each molecule was established, its molar ellipticity and ultraviolet-visible absorption spectra were measured.…”

Section: Discussionmentioning

confidence: 99%

“…The semisynthetic myoglobin was reduced to the ferro derivative with sodium dithionite and converted to the carboxy form under a steady stream of carbon monoxide. Its visible absorbance spectrum was recorded with a Cary Model 14 spectrophotometer and compared with that of the untreated carboxyferromyoglobin (Hardman et al, 1966).…”

Section: Methodsmentioning

confidence: 99%

Myoglobin semisynthesis: removal of the amino-terminal valine of sperm whale myoglobin and its subsequent reincorporation

DiMarchi¹,

Neireiter²,

Garner³

et al. 1979

Biochemistry

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“…The determination of the Fen-NOB complex formation from metmyoglobin (Mb+) and from oxidized microsomes required an approximation of the extinction coefficient for each complex. Metmyoglobin was prepared by methods previously described (Hardman et al, 1966) and gave an absorbance maximum at 412 nm. The Mb+ solution was deoxygenated under argon by successive purging and evacuation.…”

mentioning

confidence: 99%

Direct formation of complexes between cytochrome P-450 and nitrosoarenes

Jm¹,

Jf²,

Burstyn³

et al. 1986

Biochemistry

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The mechanism of the formation of the complexes between various nitrosobenzenes and cytochrome P-450 has been investigated. We have observed the formation of these complexes by a new and, as yet, undescribed route. Nitrosobenzene (NOB) itself reacts with cytochrome P-450 in the iron(III) state, in the absence of any exogenous reducing agent, to produce the iron(II)-NOB complex. Apparently, NOB is a ligand that is capable of causing the spontaneous autoreduction of the iron. The reduction of the iron may occur via ligand-induced oxidation of the axially bound thiolate of cytochrome P-450.

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Isolation of Sperm Whale Myoglobin by Low Temperature Fractionation with Ethanol and Metallic Ions

Cited by 122 publications

References 33 publications

Characterization of the reaction of methyl acetimidate with sperm whale myoglobin

Characterization of the reaction of methyl acetimidate with sperm whale myoglobin

Myoglobin semisynthesis: removal of the amino-terminal valine of sperm whale myoglobin and its subsequent reincorporation

Direct formation of complexes between cytochrome P-450 and nitrosoarenes

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