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PubMed1976

Antigoitrogenic effect of casein.

Epidemiological findings from the city of Cali, Colombia, support the hypothesis that water supply and iodine intake are not the only dietary factors which influence the magnitude of the goitre endemia. Experiments were conducted in rats to determine whether casein has a counteracting effect on the goitrogenic and antithyroid activities of methimazole (MMI) and goitrogenic water extracts (GWE) from the endemic area of the Cauca Valley. Female albino rats (Charles River, DC strain) 100-110 g initial weight, receiving 12 mug of iodine daily, were divided into three groups annd put on special diets: protein-free, 8 % casein, or 60 % casein, respectively. Each group (24 rats) was then divided into three subgroups. Subgroup one received goitrogen-free water and was used as a control. Subgroup two was administered MMI, 50 mug/day/rat. Subgroup three was given per animal a daily amount of GWE equivalent in antithyroid potency to 50 mug of MMI. At 77 days, the thyroid glands were studied for weight, 131I uptake, and 127I concentration. Animals on the protein-free diet showed significantly (P less than 0.05 - less than 0.01) larger thyroid glands per 100 g body weight and lower thyroidal 4 h 131I uptake and 127I-concentrations than rats on casein diets. These differences were significantly increased (P less than 0.01) by the administration of MMI and GWE. All the effects were completely reversed by the 60 % casein diet showing no differences between control rats and those on MMI or GWE. Rats on 8 % casein showed intermediate values between those of animals on protein-free and 60 % casein diets; differences were still present between the control as against the MMI or GWE groups, The results indicate that under these experimental conditions, a poor-protein diet impairs the thyroidal transport of iodine, decreases its concentration in the thyroid and is accompanied by an enlargement of the gland. Under these circumstances, the action of thiourea-like antithyroid agents is enhanced. The administration of protein reverses these alterations and decreases the action of such antithyroid agents. Whether the changes observed are due to a direct action of casein on the thyroid and/or to effects of malnutrition on the metabolism of antithyroid compounds remains to be determined.

Animals↗
PubMed1997

The effect of abomasal infusion of casein and recombinant somatotropin hormone injection on nitrogen balance and amino acid fluxes in portal-drained viscera and net hepatic and total splanchnic blood in Holstein steers.

Six Holstein steers, 6 mo of age, with a BW range of 180 to 200 kg were used to determine effects of abomasal casein infusion and recombinant bovine somatotropin on amino acid absorption and metabolism in the gut and liver. Catheters were positioned in the hepatic vein, the hepatic portal vein, two mesenteric veins, and a mesenteric artery. Using the same basal diet, treatments consisted of 1) basal diet only (Control), 2) abomasal casein infusion of 300 g/d (Casein), and 3) abomasal casein infusion plus daily injection of 20 mg of bovine somatotropin (ST). All steers were fed the basal diet at a rate of 24 g/kg of BW on a DM basis, in 12 equal meals at 2-h intervals. Initially, Casein and ST treatments were in a balanced cross-over design, and then all steers received the Control treatment. Casein infusion tended to increase BW gain but did not affect BW gain per unit of N intake. The ST treatment increased weight gain (P < .01) and efficiency of dietary DM (P < .01) and N (P < .01) utilization by more than 40% and increased the percentage of absorbed N retained. Nonessential, essential, and total amino acid fluxes in portal-drained viscera (PDV) and their net removal by the liver were not affected by casein infusion. However, ST tended to reduce PDV flux of essential, nonessential, and total amino acids by approximately 30%, and their removal by the liver (P < .08). As a result, ST increased, compared with casein, release of essential (P < .06) and total (P < .10) amino acids from the total splanchnic pool (TSP) into peripheral blood. The ST treatment increased the availability of absorbed amino acids to peripheral tissues, but it did not affect the amino acid profile delivered from splanchnic tissues, especially that of essential amino acids.

Abomasum↗
PubMed1998

Temporal change in skeletal muscle IGF-I mRNA abundance and nitrogen metabolism responses to abomasal casein infusion in steers.

Skeletal muscle IGF-I and alpha-actin mRNA responses to increased amino acid availability were investigated in young, rapidly growing steers. Four Holstein steers (208 kg BW) were surgically implanted with an abomasal cannula and jugular catheters and allowed 2 wk to recover. Steers were offered hourly a 43:57 forage:concentrate diet at 95% of ad libitum intake supplemented with continuous abomasal infusion of glucose (to replace 12.5% of metabolizable ad libitum energy intake) for 13 d before the start of abomasal infusion of 67 g of casein N/d. Biopsies of the liver and both semimembranosus muscles were removed and frozen in liquid N, and casein infusion was begun. Muscle biopsies were collected at 8, 16, 24, and 48 h, and on d 7 and 14. Nitrogen balance increased from 23.6 to 71.5 g/d (P < .001) within 24 h and remained elevated (mean = 58.4 g/d) during the 14 d of casein infusion. Plasma urea N increased from 4 to 9.5 mg/dL at 24 h and remained unchanged to d 14. Muscle IGF-I mRNA abundance increased to 215% of basal values at 16 h (P < .01), 244% of basal values at 24 h, and 222% of basal values at 48 h after initiation of casein infusion. Values reached a maximum of 274% of basal values on d 7 and then declined to near preinfusion levels on d 14. The IGF-I mRNA abundance was approximately 100 times higher in liver than in skeletal muscle and was not different on d 0 and 14. Although plasma IGF-I concentrations were numerically higher during the first 24 h of abomasal casein infusion, they were not significantly higher during the chronic phase of treatment. Plasma IGF binding protein (BP)-2 concentrations were higher at 16, 24, and 48 h after casein infusion was begun, but IGFBP-3 concentrations were not altered at these sampling times. Neither acute (first 24 h) nor chronic (daily) plasma insulin concentrations were altered by abomasal casein infusion. Plasma somatotropin concentrations were lower (P = .008) at 24 h of casein infusion and beyond. Results suggest that enhanced amino acid availability may modulate skeletal muscle protein synthesis and accretion through an autocrine or paracrine IGF-I influence.

Abomasum↗
PubMed2004

Pancreatic exocrine secretion and plasma concentration of some gastrointestinal hormones in response to abomasal infusion of starch hydrolyzate and/or casein.

Eight Angus steers (290 +/- 8 kg), surgically prepared with pancreatic pouch-duodenal reentrant cannulas and abomasal infusion catheters were used in a replicated 4 x 4 Latin square experiment to investigate the effects of abomasal infusion of starch hydrolyzate (SH) and/or casein on pancreatic exocrine secretion and plasma concentration of hormones. Steers were fed a basal diet of alfalfa (1.2 x NEm) in 12 equal portions daily. Abomasal infusion treatments (6-L total volume infused per day) were water (control), SH [2.7 g/(kg BW x d)], casein [0.6 g/(kg BW x d)], and SH + casein. Periods were 3 d for adaptation and 8 d of full infusion. Pancreatic juice and jugular blood samples were collected over 30-min intervals for 6 h on d 11. Weight and pH of pancreatic samples were measured, and a 10% subsample was composited and frozen until analysis of total protein and pancreatic enzyme activities. The remaining sample was returned to the duodenum. Plasma was harvested and frozen until analyzed. Pancreatic juice (67 mL/h) and protein (1.8 g/h) secretion rates were not affected by nutrient infusion. There were SH x casein interactions for all pancreatic enzyme secretions (U/h; alpha-amylase, P < 0.03; trypsin, P < 0.08; and chymotrypsin, P < 0.03) and plasma insulin concentration (P < 0.10). Secretion of pancreatic enzymes was increased by SH (trypsin) and casein (alpha-amylase, trypsin, and chymotrypsin) but not when SH + casein were infused together. Glucose (P < 0.10) and cholecystokinin octapeptide concentrations (CCK-8; P < 0.05) were increased by SH, but glucagon was decreased (P < 0.10). Casein decreased (P < 0.10) plasma CCK-8 concentrations. These data indicate that positive effects of postruminal casein on enzyme secretion were inhibited by SH, emphasizing the complexity of the regulatory mechanisms involved in dietary adaptation of pancreatic exocrine secretion. Changes in hormone concentration may not relate directly to changes in enzyme secretion.

Abomasum↗
PubMed2001

Cation-exchange purification of mutagenized bovine beta-casein expressed in transgenic mouse milk: its putative Asn68-linked glycan is heterogeneous.

Bovine beta-casein (A2 genetic variant) was mutagenized to (L70S/P71S) and expressed in transgenic mouse milk. This protein now carries the signal (N68S69S70S71) that mimics a consensus eukaryotic glycosylation signal (N-X-S/T) (3). Hypothetically this protein should be glycosylated at N68 by any eukaryotic organism producing it. This novel protein was purified from transgenic mouse milk by Mono-S cation-exchange fast protein liquid chromatography (FPLC). The novel beta-casein was separated without cross contamination from mouse caseins by using acetate buffer (pH 5.0) in the presence of 6 M urea, octyl-glucopyranoside and 2-beta-mercaptoethanol. The purified (L70S/P71S) beta-casein showed an N-linked oligosaccharide attached to Asn68 and different lectin binding profiles compared with the same protein expressed in yeast. The mouse-expressed beta-casein (L70S/P71S) was specific to Concanavalin A, wheat germ agglutinin, Erythrina cristagalli agglutinin, and Ulex europaeus, indicating its oligosaccharide structure is different in the mammary gland of mouse than the reported glycosylated beta-casein expressed in Pichia pastoris (4). In addition, the five serine residues located at amino terminus of wild type bovine beta-casein were shown to be normally phosphorylated as in native bovine beta-casein.

Animals↗
PubMed2001

Characterization of casein micelle precipitation by chitosans.

We have found that the addition of chitosan, a cationic polymer, on whole or skim milk produces destabilization and coagulation of casein micelles that takes place without changes in the milk pH or the stability of most whey proteins. The amount of lipids recovered in the chitosan-casein aggregates was similar or higher than that obtained with rennet or acid precipitation. Approximately 70% of milk Ca2+ (approximately 750 mg/L) was found in the chitosan-induced aggregates, which is 10 and 50% higher than the amounts observed with acid or rennet coagulations, respectively. Purified alpha, beta-, and kappa-caseins were extensively precipitated by different molecular weight chitosans at pH 6.8. The phosphate groups of caseins seem not to be relevant in this interaction because dephosphorylated alpha- and beta-caseins were equally precipitated with chitosans. Analysis by optical microscopy of the chitosan-casein complex reveals that the size of the aggregates increase as the molecular weight of chitosans increase. Hydrophobic and electrostatic interactions particpate in the association and coagulation of casein micelles with chitosans of different molecular weights. The phenomenon is observed over a broad range of temperature (4 to 70 degrees C) with a reduction in the concentration of chitosan needed to precipitate the caseins that parallels a reduction in the viscosity of the chitosan solutions. Taken together, the results indicate that the electrostatic interactions may contribute energetically to the association between the two biopolymers, but the hydrophobicity of the complex would be the key determinant in the overall energetics of the reaction.

Animals↗
PubMed1987

A rapid method for measuring protease activity in milk using radiolabeled casein.

A rapid means to detect the presence of protease activity in raw milk could be useful in predicting keeping ability of products made from that milk. A 30-min assay has been developed and compared with three other methods of detecting protease. Casein, [methyl-14C]-methylated-alpha was purchased from a radioisotope supplier. Concentrations of substrate from 2 to 20 nCi gave counts per minute, which increased linearly when counted with the Charm analyzer. There was not a significant difference in counting times of 10, 20, or 30 min. A mixture of sodium acetate and acetic acid precipitated nonhydrolyzed substrate with an efficiency of 97%. Comparison of the [14C] casein assay, a casein fluorescein isothiocyanate assay, trinitrobenzenesulfonic acid procedure, and the Hull procedure using protease from psychrotrophic bacteria revealed that the [14C] casein and casein fluorescein isothiocyanate methods were roughly equivalent and that the radiometric procedure was 10 times more sensitive than the trinitrobenzenesulfonic acid assay. The radiometric procedure was approximately 10(4) times more sensitive than the Hull procedure. The [14C] casein and casein fluorescein isothiocyanate methods were similar in time required, about 30 min, while the trinitrobenzenesulfonic acid assay and Hull method required about 1 h plus reagent preparation time. The [14C] casein procedure was most expensive per test; the other three were cheaper and similar to each other in cost.

Acinetobacter↗
PubMed1991

Association of kappa-casein glycosylation with milk production and composition in Holsteins.

A total of 545 milk samples were collected from 53 Holstein cows for 1 yr. On test day, morning milk yields were recorded; milk samples were analyzed for N-acetylneuraminic acid, protein, fat, casein, kappa-casein, and SCC. the relationship between the degree of glycosylation of kappa-casein and morning milk yield and composition was investigated. Data were analyzed using least squares procedures with a model that included test day, parity, stage of lactation, SCC, and phenotype for kappa-casein as fixed effects and N-acetylneuraminic acid content of kappa-casein as covariate. After adjustments were made for effects of environmental and genetic factors, the degree of glycosylation of kappa-casein was associated with morning milk yield, protein, and casein content. Milk yield increased linearly with an increase of N-acetylneuraminic content up to approximately 70 micrograms/mg of kappa-casein.

Animals↗
PubMed1996

Behavior of calcium and phosphate in artificial casein micelles.

This study investigated the behavior of Ca and phosphate in artificial casein micelles using 45Ca and 32P by means of UF, equilibrium dialysis, and gel permeation HPLC in the presence of 6 M urea. Artificial casein micelles were prepared at a casein concentration of 2.5% with 30 to 40 mM Ca, 22 to 27 mM phosphate, and 10 mM citrate using 45Ca or 32P. About 75 and 65% of colloidal 45Ca and 32P, respectively, in artificial casein micelles that were formed in the presence of 30 mM Ca and 22 mM phosphate were exchanged after dialysis of artificial casein micelles against a simulated milk ultrafiltrate at 4 degrees C for 72 h. The percentages of 45Ca and 32P in the fraction of casein aggregates that were crosslinked by micellar Ca phosphate were 26.9 and 27.6%, which decreased to about 10% after equilibrium dialysis against a simulated milk ultrafiltrate at 4 degrees C for 72 h. The proportion of exchanged colloidal 45Ca and 32P in artificial casein micelles during dialysis for 72 h were higher at 25 degrees C than at 4 degrees C, suggesting that the exchange of Ca and phosphate between the diffusible and colloidal phases depended on temperature. The results suggest that the exchange of Ca and phosphate between diffusible and colloidal phases proceeds slowly, and a portion of the Ca and phosphate in the artificial casein micelles is difficult to exchange.

Animals↗
PubMed1985

Gelation of the heat-induced complex between kappa-casein and alpha-lactalbumin.

Whey is a by-product of the manufacture of cheese from milk. The usual practice is to dispose of it, the usage of whey being not sufficiently developed, though it contains proteins of excellent quality such as beta-lactoglobulin and alpha-lactalbumin. Interaction between kappa-casein and whey protein was examined in order to form new food materials. Gelation of the heat-induced complex between kappa-casein and alpha-lactalbumin is described in this paper. alpha-Lactalbumin was easily separated from whey protein by the gel filtration technique on a Toyopearl HW-50 column at pH 6.0. Heat treatment facilitated the hydrolysis of a mixture of kappa-casein and alpha-lactalbumin by trypsin, alpha-chymotrypsin and pronase. Heat treatment at above 75 degrees C and a protein level of over 5% were needed to form gel in 35 mM phosphate buffer, pH 7.6, containing 0.4 M NaCl. The temperature was in agreement with that at which alpha-lactalbumin denatured and formed the complex with kappa-casein. Decrease of soluble protein concentration and increase of turbidity were induced with gelation. Gel was not formed when only kappa-casein or alpha-lactalbumin was heated under the appropriate conditions. It was considered that a kappa-casein-alpha-lactalbumin gel was formed from a complex of the two proteins by heat treatment. The breaking stress of kappa-casein-alpha-lactalbumin gel was less than that of kappa-casein-beta-lactoglobulin gel. If the pH was reduced to 5.8 after complex formation by the two proteins, gel was formed at a low protein concentration compared with that with no alteration of pH.

Caseins↗
PubMed1989

54Mn absorption and excretion in rats fed soy protein and casein diets.

Rats were fed diets containing either soy protein or casein and different levels of manganese, methionine, phytic acid, or arginine for 7 days and then fed test meals labeled with 2 microCi of 54Mn after an overnight fast. Retention of 54Mn in each rat was measured every other day for 21 days using a whole-body counter. Liver manganese was higher (P less than 0.0001) in soy protein-fed rats (8.8 micrograms/g) than in casein-fed rats (5.2 micrograms/g); manganese superoxide dismutase activity also was higher in soy protein-fed rats than in casein-fed rats (P less than 0.01). There was a significant interaction between manganese and protein which affected manganese absorption and biologic half-life of 54Mn. In a second experiment, rats fed soy protein-test meals retained more 54Mn (P less than 0.001) than casein-fed rats. Liver manganese (8.3 micrograms/g) in the soy protein group was also higher than that (5.7 micrograms/g) in the casein group (P less than 0.0001), but manganese superoxide dismutase activity was unaffected by protein. Supplementation with methionine increased 54Mn retention from both soy and casein diets (P less than 0.06); activity of manganese superoxide dismutase increased (P less than 0.05) but liver manganese did not change. The addition of arginine to casein diets had little effect on manganese bioavailability. Phytic acid affected neither manganese absorption nor biologic half-life in two experiments, but it depressed liver manganese in one experiment. These results suggest that neither arginine nor phytic acid was the component in soy protein which made manganese more available from soy protein diets than casein diets.

Absorption↗
PubMed2004

Validated sandwich enzyme-linked immunosorbent assay for casein and its application to retail and milk-allergic complaint foods.

Cows' milk is a commonly allergenic food. Cross-contamination of milk proteins into nondairy, kosher-pareve foods prepared on shared processing equipment can cause severe, life-threatening reactions in milk-allergic individuals. A sandwich-type enzyme-linked immunosorbent assay (ELISA; 96-well plate format) was developed for the detection of undeclared casein in foods. Rabbit anti-casein antibodies were used as the capture reagent. Food samples and standards were ground, extracted in 0.01 M phosphate-buffered saline, clarified by centrifugation, and added to the wells. Goat anti-casein antibodies were employed as the detector antibody, and the amount of antibody bound was determined with a commercial rabbit anti-goat immunoglobulin conjugated to alkaline phosphatase, with subsequent substrate reaction. Antibodies developed were specific to casein, with no cross-reaction observed with 30 foods and food ingredients. Non-milk-containing products such as fruit juices, fruit juice bars, sorbets, and dark and pareve-labeled chocolate were purchased from June 2002 through June 2003. In addition, samples allegedly causing eight milk-allergic consumer complaints were analyzed. The ELISA had a detection limit of less than 0.5 ppm of casein. The casein content in the analyzed foods ranged from less than 0.5 ppm to more than 40,000 ppm casein; undeclared casein residues were found in all of the samples implicated in allergic reactions. The levels of milk contamination in some of the other surveyed products could also be hazardous for milk-allergic consumers. This ELISA method provides a useful quality control tool for the food industry and could also be used as a validation of kosher-pareve status.

Animals↗
PubMed2000

[Study on casein allergy in children].

Casein a component of milk is used for food additives, industrial materials and drugs. However casein is known to be a main allergen in milk allergy. Recently several cases of anaphylaxis to antibiotics including casein have been reported. In this study we investigated casein allergy in milk allergy. 6 out of 8 patients who were positive for milk RAST were also positive for casein RAST. In these positive cases only 3 out of 6 patients had some allergic symptoms after taking antibiotics. In 3 patients DLST was also positive to casein. There was one patient who was positive in DLST without any symptoms after taking the same antibiotics. It is needed to pay attention to casein allergy when giving the medication which includes casein.

Adolescent↗
PubMed2004

[Effect of casein and protamine on the enzymatic degradation and the orally hypoglycemic action of insulin].

AIM: To study the protection of casein and protamine against degradation of insulin (INS) by proteolysis enzymes and the effect of these two kinds of protein on the hypoglycemic action of INS solution and enteric-microspheres after administrated orally to rats. METHODS: HPLC was used to determine the remained INS in the solution of alpha-chymotrypsin and trypsin with or without casein or protamine; INS solution and enteric-microspheres were prepared and adiministrated orally to rats together with the absorption enhancer sodium N-[8-(2-hydroxybenzoyl) amino] caprylate (SNAC). At the same time, casein or protamine or both of these two kinds of protein were administrated together in order to study their influence on the hypoglycemic effect of INS and microspheres. RESULTS: Casein had a good protection against degradation of INS by alpha-chymotrypsin, but protamine had no protection effect. However, the degradation of INS by trypsin is concerned, the protection effect of protamine on INS was better that of casein. Both of protamine and casein can increase the hypoglycemic effect of INS solution and enteric-microspheres. Co-administrated these two kinds of protein had a better effect. In addition, co-administrated with SNAC, casein and protamine, INS enteric-microspheres had a longer and more potent hypoglycemic effect than that of the solution. CONCLUSION: Casein and protamine can increase the stability of INS in the intestinal fluid by the mechanism of competition and combine with proteolysis enzymes, which will benefit to INS oral administration.

Administration, Oral↗
PubMed1991

[Genotyping the bovine kappa-casein locus using polymerase chain reaction].

Polymerase chain reaction (PCR) was used for detecting kappa-casein (kappa-casein) genotype in the synthetic breed (Jersey x Black and White x Holstein-Friesian) dairy cattle. The amplified 228 bp fragment includes a region, where relevant mutations lead to both the appearance of different kappa-casein alleles associated with amino acid substitutions and the appearance of new TaqI and HindIII restriction sites in ae-casein B gene. The specificity of the kappa-casein gene fragment amplification was supported by restriction analysis and Southern blot hybridization. Digestion of amplified fragment with endonucleases PstI, HindIII and/or TaqI allows detection of AA-, AB- and BB genotypes of kappa-casein. A total of 32 animals with known (18 samples) and unknown (14 samples) kappa-casein phenotypes were tested using PCR and blot hybridization. In all known cases the detected genotype confirmed the phenotype. Frequencies of the B allele and of the AB genotype in the breeding population are rather high (53.1 +/- 8.8 and 43.7%, respectively). The possibility of effective use of the PCR analysis for genotyping kappa-casein locus in bulls and their offspring has been shown. The advantages of the PCR method in large breeding programs and linkage analysis have been discussed.

Animals↗
PubMed1988

Recovery of phenytoin from solutions of caseinate salts and calcium chloride.

The recovery of phenytoin from solutions of intact protein components of enteral nutrition products was studied. Diluted phenytoin oral suspension was added to 10 1-mL samples of solutions of sodium caseinate, calcium caseinate, a mixture of sodium and calcium caseinates, and calcium chloride as well as to 10 1-mL samples of a distilled water control to produce theoretical concentrations of 10 micrograms/mL. The samples were filtered using an ultrafiltration technique and assayed for phenytoin concentration by high-performance liquid chromatography. The mean concentration of phenytoin in the filtrates of the test samples was significantly lower than the mean concentration in the control samples. The recovery of phenytoin from sodium caseinate solutions was significantly lower than that from solutions of calcium caseinate or calcium chloride. There was no significant difference between recovery of phenytoin from sodium caseinate solutions and that from solutions of sodium and calcium caseinates. Decreased phenytoin absorption from enteral nutrient solutions in vitro may be associated with the presence of caseinate salts and calcium chloride. However, this relationship cannot be extrapolated to in vivo absorption.

Calcium Chloride↗
PubMed1985

Alteration of aminoacyl-tRNA synthetase activities by phosphorylation with casein kinase I.

The phosphorylation of a highly purified aminoacyl-tRNA synthetase complex from rabbit reticulocytes by the cyclic nucleotide-independent protein kinase, casein kinase I, has been examined, and the effects of phosphorylation on the synthetase activities were determined. The synthetase complex, purified as described (Kellermann, O., Tonetti, H., Brevet, A., Mirande, M., Pailliez, J.-P., and Waller, J.-P. (1982) J. Biol. Chem. 257, 11041-11048), contains seven aminoacyl-tRNA synthetases and four unidentified proteins and is free of endogenous protein kinase activity. Incubation of the complex with casein kinase I in the presence of ATP results in the phosphorylation of four synthetases, namely, glutamyl-, isoleucyl-, methionyl-, and lysyl-tRNA synthetases. Phosphorylation by casein kinase I alters binding of the aminoacyl-tRNA synthetase complex to tRNA-Sepharose. The phosphorylated synthetase complex elutes from tRNA-Sepharose at 190 mM NaCl, while the nonphosphorylated complex elutes at 275 mM NaCl. Phosphorylation by casein kinase I results in a significant inhibition of aminoacylation by the glutamyl-, isoleucyl-, methionyl-, and lysyl-tRNA synthetases; the activities of the nonphosphorylated synthetases remain unchanged. These data indicate that phosphorylation of aminoacyl-tRNA synthetases in the high molecular weight complex alters the activities of these enzymes. One of the unidentified proteins present in the complex (Mr 37,000) is also highly phosphorylated by casein kinase I. From a comparison of the properties and phosphopeptide pattern of this protein with that of casein kinase I, it appears that the Mr 37,000 protein in the synthetase complex is an inactive form of casein kinase I. This observation provides further evidence for a physiological role for casein kinase I in regulating synthetase activities.

Amino Acyl-tRNA Synthetases↗
PubMed1982

Phosphorylation of glycogen synthase by cyclic AMP-independent casein kinase-2 from rabbit skeletal muscle.

Cyclic AMP-independent casein kinase-2 from rabbit skeletal muscle has been extensively purified by procedures including phosphocellulose, Bio-Gel A-1.5m, casein-Sepharose, and DEAE-cellulose column chromatographies. The casein kinase and glycogen synthase kinase activities are co-purified throughout the purification. The casein kinase-2 has Mr approximately equal to 135,000 as determined by glycerol density gradient centrifugation and by gel filtration. Analysis of the purified kinase by gel electrophoresis in the presence of sodium dodecyl sulfate reveals the presence of two major protein bands having Mr = 42,000 and 27,000 in a ratio of 0.85:1. The kinase phosphorylates glycogen synthase, casein, and phosvitin either in the presence of ATP or GTP; however, the Km values for GTP are slightly higher than those of ATP. The activity of this kinase is inhibited by heparin but is not affected by the addition of cyclic AMP, heat-stable inhibitor of cyclic AMP-dependent protein kinase, Ca2+, ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, or the combination of Ca2+ and calmodulin. The phosphorylation of the synthase by the kinase results in the incorporation of approximately 0.8 mol of PO4/subunit and a reduction in the synthase activity ratio from 0.82 to 0.6. The extent of phosphorylation of the synthase catalyzed by casein kinase-2 is similar to that by phosphorylase kinase; however, the sites phosphorylated by these two kinases are different. The relationships of casein kinase-2 to the cyclic AMP-independent synthase kinases that have been described by other laboratories and to the casein kinases isolated from other tissues are discussed.

Animals↗