Influence of fatty acids and related anions on the azorubin-binding capacity of serum albumin

Westphal, Ulrich; Stets, John F.; Priest, Stanley G.

doi:10.1016/0003-9861(53)90141-6

Cited by 16 publications

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“…They were dialyzed in a mechanically stirred dialyzer (5) The various dialysates of the sera were acidified and extracted several times with petroleum ether. The dry residues of the petroleum-ether phase were combined with human serum albumin as described before (1). For the direct extractions of normal and low-ABC sera, alcohol-ether mixtures (3: 1) were used at normal and elevated temperatures; the lipid material was also combined with albumin.…”

Section: Methodsmentioning

confidence: 99%

Increased Electrophoretic Mobility of Albumin in the Serum of Rats Subjected to Carbon-Tetrachloride Treatment or to Tourniquet Shock

Westphal¹,

Priest²

1955

J. Clin. Invest.

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It has been previously reported (1) that binding of small quantities of certain fatty acids to serum albumin causes a decrease of its azorubin-binding capacity (ABC). It was assumed then that the low ABC of serum albumin observed in clinical cases (2) or in experimental animals (3, 4) was produced by a firm binding, in vivo, of increased amounts of certain anions, possibly higher fatty acids.If this explanation were correct, a negative increment in the net charge of the albumin molecule would be expected. The electrophoretic mobilities of albumin were therefore determined in sera of rats subjected to carbon-tetrachloride poisoning and to tourniquet shock since these conditions are known to lower the ABC of serum albumin (3). Studies on the influence of dialysis on the ABC of sera and other protein solutions were included as supporting data. EXPERIMENTALAnimal experiments. Male Sprague-Dawley rats were given i.p. injections of 0.5 ml. CC4 per Kg. body weight every second day for six days and twice this volume of a 1 + 1 mixture of CCI4 and ethanol on the eighth day. The blood was obtained on the tenth day. Other details have been given previously (3). The average weight of the rats declined from 336 gm. to 271 gm. under the injections of CCI4. Tourniquet shock was produced by applying rubber bands to both hind legs (3). The average weight of the animals at the time of sacrificing was 269 gm. The blood of all experimental animals was drawn from the heart and the serum prepared as described (3). The average non-protein nitrogen concentration in the sera listed in Table I was found to be 72 mg. per cent, indicating a severe state of shock. The various samples of serum for the normal, CCL-treated and tourniquet-shock rats were obtained from a total of 88, 163, and 148 animals, respectively. Chemical procedures. Total protein and non-protein nitrogen were determined by a micro-Kjeldahl procedure as outlined previously (3).Electrophoretic analysis. The sera were dialyzed against sodium phosphate buffer of pH 7.7, iA= 0.1, for a total of 22 hr. at 20 C. in a mechanically stirred dialyzer (5) with six changes of buffer. The diluted serum solution was used for the measurement of the pH (glass electrode, room temperature) and resistivity. The electrophoretic analyses were done in a standard size electrophoresis apparatus 1 using the long analytical cell. The separations were mostly run for 120 minutes at + 0.40 C. and 24 mA. The conductivity was measured in a Shedlovsky cell equilibrated at + 0.40 C.The evaluation of the electrophoretic patterns was done as in earlier studies (3), using the descending boundaries only. The procedure of Longsworth and MacInnes (6) was applied to determine the distance traveled by albumin. As a check of the accuracy of the electrophoretic analysis, the mobility of crystalline bovine serum albumin (Armour) was determined in a 1 per cent solution in glycine sodium hydroxide buffer, IA= 0.2 at pH 8.4. The value obtained (-5.79 X 0I cm.' sec.:1 volt71 at pH 8.35) was in agreement with the mobil...

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Section: Methodsmentioning

confidence: 99%