Note on spectrophotometric determination of proteins in dilute solutions

Murphy, Joseph B.; Kies, Marian W.

doi:10.1016/0006-3002(60)91464-5

Cited by 572 publications

(126 citation statements)

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“…When radial immunodiffusion was performed with isolated mitochondria, the same ratio for spf CRM/control CRM was found [ 1,2]. This indicates that mature intra-mitochondrial OTCase is increased in spf mice and rules out the possibility of a mitochondrial membrane defect in the transport of spf pre-OTCase from the cytosol.…”

Section: Determination Of the Amount Ofmentioning

confidence: 85%

“…The high activity of spf OTCase at pH 9 observed in crude homogenates appears to be the result of two kinds of events: [1] (1) (2) An enhancement in the amount of OTCase protein in liver mitochondria as demonstrated by immunological measurements. It must therefore be assumed that a mechanism enhances the amount of mutant OTCase which tends to counter-balance the decrease in its catalytic activity at the physiological pH of the cell.…”

Section: Conclusion Referencesmentioning

confidence: 99%

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Increase of ornithine transcarbamylase protein in sparse‐fur mice with ortnithine transcarbamylase deficiency

et al. 1981

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Section: Determination Of the Amount Ofmentioning

confidence: 85%

Section: Conclusion Referencesmentioning

confidence: 99%

Increase of ornithine transcarbamylase protein in sparse‐fur mice with ortnithine transcarbamylase deficiency

et al. 1981

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“…During this period the animals were placed separately in siliconized metabolism cages (CLEA CT-10), and given NaCI solution in fixed concentration (15,40,77 or 120 mM) and a low sodium diet. The latter was produced in our laboratory according to the components of the CLEA low sodium diet (Mixture No.…”

Section: Methodsmentioning

confidence: 99%

Natriuresis of Fasting in Intact and Adrenalectomized Rats

Nohno

Hayashi

Murayama

1977

Japanese Journal of Pharmacology

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Abstract-Experimentswere carried out in an attempt to elucidate the mechanism of fasting-induced natriuresis in conscious rats. Female animals were given a low sodium diet and saline in a fixed concentration for at least three days, and deprived of food thereafter. The sodium balance significantly shifted to negative independently of potassium supply in intact rats. Such was also observed in the dexamethasone replaced, adrenalectomized rats and was not affected by further administration of aldosterone.In addition, the diuretic effect of insulin in fasted intact rats was not evident in the fed diabetic rats in which diabetes had been induced by streptozotocin. Such findings suggested participation of factors other than insulin. The natriuresis of fasting in intact rats appears to involve two factors, one of which is independent of the sodium intake level. Dependence on the sodium intake level may be derived from alteration of the solute diuresis-like effect of drinking saline when animals are fasted. These results suggest that neither aldosterone nor insulin is a major causal factor involved in fasting-induced natriuresis.In man and rabbit, many workers have shown that fasting produced natriuresis, and such was arrested by carbohydrate feeding (1-8). Previous reports indicated that in rats also, food deprivation resulted in an increase in sodium excretion or a negative shift of sodium balance. Several hormones regulating the glucose metabolism may be involved in the occurrence of sodium imbalance during the early stage of fasting, however the mecha nism remains obscure.The antidiuretic effect of insulin (9, 10) has been attributed to fasting-induced natriuresis and anti -natriuresis by subsequent glucose feeding. In a preliminary report from this laboratory it was confirmed that the negative balance of sodium in fasted rats was not improved but rather enhanced by the administration of insulin (11). Since glucagon has a natriuretic effect in man and the dog (12)(13)(14), it is suggested that glucagon may contribute either directly or indirectly to the natriuretic effect of fasting. Moreover, well-known factors, such as aldosterone and antidiuretic hormone, may be implicated in the sodium imbalance.The present series of studies on the fasting-produced natruiresis was undertaken to determine: (a) The role of sodium and potassium loading in intact fasting rats; (b) the effect of aldosterone in glucocorticoid-replaced adrenalectomized rats; and (c) the effect of insulin in intact or diabetic rats. MATERIALS AND METHODSFemale Wistar rats weighing between 190 and 310 g were maintained on a regular diet

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“…Immunoblot analysis was performed using a sheep-anti-human Cu,Zn-SOD antibody (Calbiochem). Protein concentration was determined by the method of [ 10]. The identities of the purified wild-type and D90A proteins were verified by electrospray mass spectrometry (VG, Qurtto).…”

Section: Analysis Of Recombinant Cu Zn-sodmentioning

confidence: 99%

The free radical‐generating function of a familial amyotrophic lateral sclerosis‐associated D90A Cu,Zn‐superoxide dismutase mutant

Kim¹,

Eum²,

Kwon³

et al. 1998

IUBMB Life

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SUMMARYThe free radical-generating functions of the D90A Cu,Zn-superoxide dismutase (SOD) associated with Swedish familial amyotrophic lateral sclerosis (FALS) patients are investigated. The results show that both the wild-type and mutant enzymes have identical dismutase activity, while the free radical-generating activity of the D90A mutant is enhanced relative to that of the wild-type enzyme. The studies suggest that the active channel of the D90A mutant is larger than that of the wild-type enzyme. A higher free radical-generating activity of the mutant enzyme led to the release of copper ions from the damaged protein. The generation of strand breaks in plasmid DNA was enhanced more effectively by the D90A mutant Cu,Zn-SOD than by the wildtype enzyme. The results suggest that the pathology of FALS may be attributed to oxidative damage caused by the gain-of-function of FALS Cu,Zn-SOD mutant.Key words: copper,zin-superoxide dismutase, mutant, hydroxyl radical, copper. 1191All rights of reproduction in any form reserved, Vol. 46, No. 6, 1998 BIOCHEMISTRY and MOLECULAR BIOLOGY INTERNATIONALAndersen et al. [7] reported a Cu,Zn-SOD exon 4 mutation (D90A) with novel features in Swedish FALS patients and the disease has only been observed in patients homozygous for this mutation, whereas heterozygotes remained unaffected. Furthermore, there was essentially no reduction in erythrocyte Cu,Zn-SOD dismutation activity. Thus, [7] suggested that this Cu,Zn-SOD mutation caused ALS by a gain-of-function rather than by the loss of dismutation activity.In view of these considerations, the present work represents an investigation of the free radical-generating functions of the D90A mutant and the oxidative damage to DNA, as a first attempt to establish the extent of any correlation between the gain-of-function and the pathology of FALS. MATERIALS AND METHODS Expression and purification of wild-type and mutated human Cu, Zn-SOD

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Note on spectrophotometric determination of proteins in dilute solutions

Cited by 572 publications

References 4 publications

Increase of ornithine transcarbamylase protein in sparse‐fur mice with ortnithine transcarbamylase deficiency

Increase of ornithine transcarbamylase protein in sparse‐fur mice with ortnithine transcarbamylase deficiency

Natriuresis of Fasting in Intact and Adrenalectomized Rats

The free radical‐generating function of a familial amyotrophic lateral sclerosis‐associated D90A Cu,Zn‐superoxide dismutase mutant

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