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PubMed2004

Nitrogen balance in lambs fed low-quality brome hay and infused with differing proportions of casein in the rumen and abomasum.

Twenty wether lambs (46 +/- 2 kg) fitted with ruminal and abomasal infusion catheters were used in a completely randomized design to determine the effects of differing proportions of ruminal and abomasal casein infusion on N balance in lambs fed low-quality brome hay (0.8% N, DM basis) for ad libitum intake. Wethers were infused with 0 (control) or 10.7 g/d of N from casein with ratios of ruminal:abomasal infusion of 100:0 (100R:0A), 67:33 (67R:33A), 33:67 (33R:67A), or 0:100% (0R:100A), respectively, over a 12-d period. Total N supply (hay N intake + N from casein infusion) was greater (P = 0.001) in lambs receiving casein infusion than in controls. Urinary N excretion (g/d) was greater (P = 0.001) in lambs receiving casein infusion than in controls. Urinary N excretion decreased as casein infusion was shifted from 100R:0A to 33R:67A and then slightly increased in lambs receiving 0R:100A (quadratic, P = 0.02). Total N excretion was greater (P = 0.001) in lambs receiving casein infusion than in controls and decreased linearly (P = 0.005) as casein infusion was shifted to the abomasum. Retained N (g/d, % of N intake, and % of digested N) was greater (P = 0.001) in lambs receiving casein than in controls. Retained N increased as infusion was shifted from 100R:0A to 33R:67A and then slightly decreased in lambs receiving 0R: 100A (quadratic, P < 0.07). Based on regression analysis, the predicted optimum proportion of casein infusion to maximize N retention was 68% into the abomasum. The regression suggests that supplementation with undegradable intake protein had an additional benefit over supplementation with ruminally degradable intake protein (100R:0A) and that changing the percentage of ruminally undegradable intake protein in supplemental protein from 33 to 100% resulted in minimal differences in N retention. Apparent N, DM, OM, and energy digestibility (% of intake) was greater (P < 0.03) in lambs infused with casein than controls but did not differ among casein infusion groups. These data suggest that feeding protein supplements containing a portion (greater than 0%) of the crude protein as ruminally undegradable intake protein, as compared to 100% ruminally degradable intake protein, to lambs consuming low-quality forage increases N retention and the efficiency of N utilization without influencing total-tract nutrient digestion.

Abomasum↗
PubMed2001

Proteolysis of beta-casein as a marker of Grana Padano cheese ripening.

Proteolysis has a critical role in defining the typical organoleptic characteristics of Grana Padano, a well-known Italian cheese. During the ripening process, hydrolysis of beta-casein produces different fragments, the most abundant and widely studied of which are gamma-caseins, three polypeptides containing the HOOC-terminal portion of beta-casein. By sodium dodecyl sulfate-PAGE and a specific anti-beta-casein monoclonal antibody, two beta-casein-derived bands were identified in Grana Padano cheese: betaa and betab. Thanks to the identification of the amino acid sequences, it was shown that: a) betaa contains gamma1-casein [beta-casein (29-209)] and the correlated peptide [beta-casein (30-209)]; b) betab contains gamma2-casein [beta-casein (106-209)] and gamma3-casein [beta-casein (108-209)]. The production of betaa and betab by the three enzymes most involved in cheese proteolysis (pepsin, chymosin, and plasmin) was evaluated by performing in vitro digestions. A significant correlation between abundance of some polypeptides and ripening process was shown.

Amino Acid Sequence↗
PubMed2005

Relationship between physical properties of casein micelles and rheology of skim milk concentrate.

The properties of casein micelles in milk concentrates are of interest for the use of ultrafiltered (UF) skim milk concentrates in dairy products, and for the general understanding of colloidal stability and behavior of the casein micelle. The rheological behavior of UF skim milk concentrate with a casein concentration of 19.5% (wt/wt) was investigated at different pH and NaCl concentrations by analyzing flow viscometry and small amplitude oscillatory shear measurements. Viscometric flow curves were fitted to the Carreau-Yasuda model with the aim of determining values for the viscosity at infinite high shear rates and thereby estimate the voluminosity of the casein micelles (nu(casein)) in the UF concentrate. The voluminosity of the casein micelles increased with addition of NaCl and decreased when pH was decreased from 6.5 to 5.5. At pH 5.2, nu(casein) increased because of acid-induced aggregation of the casein micelles. The changes in nu(casein) could be interpreted from transmission electron microscopy of freeze-fractured samples of the UF concentrate and partly from dynamic light scattering measurements. Altered interactions between casein micelles due to different pH and NaCl concentrations are proposed to occur due to collapse of the kappa-casein layer, changed ionic strength, and altered distance between casein micelles.

Animals↗
PubMed1983

Tissue- and cell-specific casein gene expression. II. Relationship to site-specific DNA methylation.

The relationship between DNA methylation and the expression of the gamma- and beta-casein genes was investigated in both expressing and nonexpressing tissues and in isolated tumor cell subpopulations displaying differential casein gene expression. MspI/HpaII digestions of DNA isolated from liver, a totally nonexpressing tissue, indicated that specific sites of hypermethylation existed in these genes as compared to the DNA isolated from casein-producing lactating mammary gland. The positions of these sites were mapped in the gamma-casein gene by comparing total genomic DNA Southern blots to the restriction digests of several overlapping phage clones constituting the gamma-casein gene. In contrast, the methylation status of the HhaI sites in the gamma-casein gene was found to be invariant regardless of the expression status of the gene. The inverse correlation between the hypermethylation of certain MspI/HpaII restriction sites in the casein genes and their potential expressibility was further substantiated by studies in 7,12-dimethylbenz(a)anthracene- and N-nitrosomethylurea-induced mammary carcinomas, which have an attenuated casein gene expression, and in cell subpopulations isolated from the 7,12-dimethylbenz(a)-anthracene tumor which were either depleted or enriched in casein-producing cells. Analysis of total tumor DNAs indicated that the casein genes were hypermethylated at the same sites observed in liver. However, a very faint hybridization signal was observed in the HpaII digests, suggesting cell-specific methylation differences. We have confirmed the hypomethylation of at least two of these MspI/HpaII sites within the subpopulation containing the casein-producing cells at a level consistent with the relative enrichment in that fraction. These results demonstrate differential site-specific casein gene methylation not only between tissues but also between cell subpopulations within a single tissue.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1997

Monoclonal and polyclonal antibodies against casein components of cow milk for evaluation of residual antigenic activity in 'hypoallergenic' infant formulas.

BACKGROUND: Hydrolysed casein and whey protein formulas have been developed with the aim of preventing sensitization in infants at high risk of cow milk allergy. Subsequently these products have also been used for treatment of children with cow milk allergy. However, severe reactions have occurred in some allergic infants fed with these formulas raising doubts about their absolute safety and suggest the need for developing in vitro techniques for detection of eventual residual allergenic activity in such preparations. OBJECTIVES: Our purpose was to evaluate the usefulness of monoclonal and polyclonal antibodies against casein components (alpha, beta and kappa casein) as reagents for the detection of the residual antigenic activity of casein components in several hydrolysed formulas. METHODS: The monoclonal and polyclonal antibodies were produced according to standard procedures by immunizing female Balb/c mice with casein fraction (a mixture of alpha, beta and kappa casein). ELISA assays were developed to test the specificity of the antibodies and to detect and evaluate the amount of residual antigenic activity of the casein components in hydrolysed formulas. RESULTS: Use of polyclonal antiserum specific for casein allowed detection of residual antigenic activity of casein components in all partial hydrolysates and in the two extensive whey protein hydrolysates in the amounts ranging from 0.05 to 0.67% of total protein. No such activity was detectable in either the two extensive casein hydrolysates tested or the aminoacid based formula. The polyclonal antiserum proved to be more suitable than monoclonals for detecting residual antigenic activity in the hydrolysates. The monoclonal antibodies were directed against epitopes expressed on different casein components. CONCLUSIONS: In this study the ELISA inhibition assay with polyclonal antibodies specific for casein components of cow milk proved to be a sensitive method for estimating residual antigenicity in the hydrolysed formulas commercially available for infants with cow milk allergy suggesting their potential application for the quality control of hypoallergenic infant formulas.

Animals↗
PubMedDate not supplied

Indirect and direct determination of the casein content of milk by Kjeldahl nitrogen analysis: collaborative study.

The classic method for determination of milk casein is based on precipitation of casein at pH 4.6. Precipitated milk casein is removed by filtration and the nitrogen content of either the precipitate (direct casein method) or filtrate (noncasein nitrogen; NCN) is determined by Kjeldahl analysis. For the indirect casein method, milk total nitrogen (TN; Method 991.20) is also determined and casein is calculated as TN minus NCN. Ten laboratories tested 9 pairs of blind duplicate raw milk materials with a casein range of 2.42-3.05% by both the direct and indirect casein methods. Statistical performance expressed in protein equivalents (nitrogen x 6.38) with invalid and outlier data removed was as follows: NCN method (wt%), mean = 0.762, sr = 0.010, sR = 0.016, repeatability relative standard deviation (RSDr) = 1.287%, reproducibility relative standard deviation (RSDr) = 2.146%; indirect casein method (wt%), mean = 2.585, repeatability = 0.015, reproducibility = 0.022, RSDr = 0.560%, RSDR = 0.841; direct casein method (wt%), mean = 2.575, sr = 0.015, sR = 0.025, RSDr = 0.597%, RSDR = 0.988%. Method performance was acceptable and comparable to similar Kjeldahl methods for determining nitrogen content of milk (Methods 991.20, 991.21, 991.22, 991.23). The direct casein, indirect casein, and noncasein nitrogen methods have been adopted by AOAC INTERNATIONAL.

Animals↗
PubMed1998

Casein kinase II activity of buffalo sperm chromatin.

Two cyclic AMP-independent protein kinase activities have been found associated with buffalo sperm chromatin: a histone kinase highly specific for arginine-rich histone was reported recently (Mudgal et al., 1997: Arch Andrology 38:191-199) and a casein kinase II is described here. Casein kinase activity was solubilized with 0.35 M NaCl, which extracted 90% of the initial enzyme activity associated with buffalo sperm chromatin. Of the two acidic proteins tested, casein was preferred substrate over phosvitin. Among the casein fractions, the order of preference for casein kinase was beta-casein > alpha-casein > casein. Cyclic AMP at concentrations up to 50 microM had no effect on the phosphorylation of casein. Phosphoamino acid analysis using casein as the substrate showed threonine to be the acceptor amino acid for phosphoester link. Phosphorylation specificity was determined by phosphorylating buffalo beta-casein followed by the preparation of tryptic peptides and identification of amino acid residue phosphorylated. Threonine residue at position 41 having clusters of acidic amino acid residues (Thr. Glu. Asp. Glu) C-terminal to it was phosphorylated, a phosphorylation specificity akin to CKII. It is thought that phosphorylation of histones decreases their association with DNA and probably makes the DNA more available for replication, while phosphorylation of nonhistone proteins modifies their interaction with histones, allowing control of template activity. Two protein kinases found in buffalo sperm chromatin may perform a similar function.

Amino Acid Sequence↗
PubMedDate not supplied

Two-dimensional electrophoresis and computer imaging: quantitation of human milk casein.

Because human casein does not precipitate from milk at its isoelectric point as does bovine casein, there is no easy method of quantitation. Casein represents only approximately 30% of the protein fraction in human milk, and the complex methods necessary for isolation cannot be used easily with small samples in a survey of a large number of mothers. Two-dimensional electrophoresis coupled with computer imaging has the potential to compare and quantitate proteins expeditiously using a small sample size. IsoDalt, a denaturing methodology, separates the casein micelle into its component parts, beta-casein, kappa-casein, parakappa-casein and casomorphins. Identification of these spots was made by immunoassay of a Western blot with monoclonal anti-human casein. Two spots at 24 kDa and 26 kDa, thought to be phosphorylated isomers of beta casein, were selected for quantitation. Milk samples from 20 mothers, 8 weeks post partum, were run on two-dimensional (2-D) gels; a slide was taken of each silverstained gel with a Kodak control strip; the slide was scanned into powerMac Photoshop 3 with a Polaroid-Sprintscan; spots were isolated using "threshold", "mask" with IPTK (Imaging Processing Tool Kit, Reindeer Games) a Photoshop plug-in, and transferred to the NIH-Image program. Using an NIH-Image gel macro (Thomas Seebacher), the area and integrated density of the spots were measured. The Kodak control scale provided calibration and conversion to OD units. Visual scanning of the gels and computer units indicated a wide range of concentrations. To understand the range in units of weight, a standard was generated using bovine alpha casein (Sigma). Measurements will be used in a statistical program, Statview (Abacus), in an attempt to correlate information from a questionaire with casein concentration.

Animals↗
PubMed2001

Selective fortification of lysinoalanine, fructoselysine and N epsilon-carboxymethyllysine in casein model systems.

In the present study, a promising strategy to study nutritional effects of selected chemical reaction products formed in heat treated protein containing foods is addressed. In due course, a selective fortification of different marker compounds for lysine damage in casein-sugar mixtures was performed to provide model systems being applicable to investigate biological effects of the cross-link lysinoalanine (LAL), the MRPs fructoselysine (FL) and N epsilon-carboxymethyllysine (CML) in a casein-linked preparation. The three different model proteins, casein-LAL, casein-FL and casein-CML were prepared by heating casein either in strong alkaline conditions at 105 degrees C for 1 h, in the presence of glucose at 65 degrees C for 68 h, or in the presence of glyoxylic acid at 37 degrees C for 19 h. Finally, the degree of lysine modification achieved was 39%, 75% and 55% for the casein-LAL, casein-FL and casein-CML, respectively. The calculation of lysine recovery and the respective analysis of each single modified casein (LAL-, FL- and CML-MP) for the selected fortified compound and each other compound vice versa proved that the individual procedure provides a specific fortification for LAL, FL and CML, respectively. The modified proteins are suitable as reference model proteins to be investigated for specific biological and toxicological effects of casein-linked LAL, FL and CML.

Caseins↗
PubMed2002

Controlled enzymatic removal of damaging casein layers on medieval wall paintings.

A new, gentle enzymatic method was developed for a controlled removal of casein layers from medieval wall paintings. These casein layers were applied over the last 60 years on wall paintings in order to decrease substantial damage due to a peeling off of the frescoes from the roughcast surface due to environmental effects. However, due to the aging of the casein layers (at 40-50 years), a more drastic peeling occurred and the danger of total destruction of the wall paintings is severe. Thus, screening was performed to find the most suitable enzyme for casein digestion. Alcalase 2.5 DX L was the most appropriate enzyme for an effective proteolysis reaction. The enzyme was immobilized on functionalized cellulose membrane. A membrane pad system with immobilized enzymes was developed which could be pressed on the casein layers on the wall painting. A controlled removal of the casein layers by proteolytic digestion was observed and it was possible to continuously wash off the hydrolyzed casein fragments from the wall painting surface by an aqueous carbonate buffer flowing through the membrane pad. The removal and the digestion was monitored by reverse HPLC. Additionally, an on-line monitoring system was set up in order to continuously follow the casein layer removal and the digestion procedure directly on the wall painting. This technique is based on noninvasive 2D-fluorescence monitoring. Optical fiber systems were used to continuously monitor the fluorescence intensity of casein-bound tryptophan. The off-line data were verified with the on-line 2D-fluorescence data. Based on the scientific result an appropriate technique for the controlled enzymatic removal of damaging casein layers on the surface of medieval wall paintings using immobilized enzyme is now available. It is now applied to remove such casein layers from medieval wall paintings in the Allerheiligen-Kapelle Cloister, Wienhausen, Germany, and the St. Alexander Kirche, Wildeshausen, Germany.

Caseins↗
PubMed1981

The effect of different proportions of casein in semipurified diets on the concentration of serum cholesterol and the lipoprotein composition in rabbits.

The effect of different proportions of casein in semipurified diets on the concentration of serum cholesterol and the lipoprotein composition was studied in rabbits. Low-casein diets (10% w/w) resulted in serum cholesterol levels and growth rates that were lower than high-casein diets (40%). An intermediate proportion of casein (20%) produced intermediate concentrations of serum cholesterol, but only minor differences in food intake and weight gain, compared with the high-casein group. In the animals with the highest values of total serum cholesterol (the 40% casein group), most of the serum cholesterol was transported in the very low density lipoproteins, whereas with moderate hypercholesterolemia (the 20% casein group), the low density lipoproteins were the main carriers of cholesterol. Elevation in lipoprotein cholesterol was associated in all groups with an increased ratio of cholesterol to protein, suggesting the formation of particles relatively rich in cholesterol. When the rabbits on the diet containing 10% casein were subsequently transferred to the 40% casein diet, a steep increase in the level of serum cholesterol occurred. Conversely, switching the rabbits on the 40% casein diet to the 10% casein diet resulted in a decrease in the level of serum cholesterol.

Animals↗
PubMed1994

Effects of low casein and fish oil on hyperlipidemia and proteinuria in nephritic rats.

The effects of amino acid-fortified low casein and fish oil (FO) diets on hyperlipidemia and proteinuria were studied in rats with nephrotoxic serum nephritis. After an antiserum injection, rats were maintained for 14 d on four different experimental diets: a 20% casein diet containing corn oil (CO) or FO, or an 8% casein diet supplemented with cystine plus threonine containing CO or FO. The 8% casein diets reduced urinary protein excretion in nephritic rats without inducing severe growth retardation or fatty liver compared with the basal 20% casein diets. Both the 8% casein diet and the FO diet decreased serum cholesterol, triglyceride and phospholipid levels in nephritic rats, and nonesterified fatty acid levels were decreased by FO feeding. In nephritic animals, hepatic cholesterol synthesis was decreased by the 8% casein diets compared with the 20% casein diets, and tended to be reduced by FO feeding between groups at the same casein levels. No effect of diet was observed on fatty acid synthesis among the nephritic rats. FO administration to the nephritic animals suppressed fecal steroid excretion. While lipoprotein lipase activity was unchanged among the nephritic rats, hepatic triglyceride lipase activity was reduced by either the 8% casein or FO diet. The results suggest that the hypolipidemic action of low casein diets may, at least in part, be due to reduced hepatic cholesterol synthesis and suppressed triglyceride secretion from the liver. They also suggest that the hypolipidemic action of FO may, at least in part, be due to reduced hepatic cholesterol synthesis and decreased fatty acid mobilization from peripheral adipose tissue.

Animal Feed↗
PubMed1982

Purification and characterization of the cytoplasmic casein kinase I from rat liver.

Two cyclic nucleotide-independent protein kinases, which preferentially utilize casein and phosvitin as substrates, exist in rat liver. In contrast to cytosol the "light' form of these enzymes was predominant in the "microsomal extract'. This form (30 000-40 000 daltons, casein kinase I) was separated from the "heavy' form (130 000 daltons, casein kinase II) by gel filtration. This enzyme was then purified by successive chromatography on carboxymethyl-Sephadex, phosvitin-Sepharose and hydroxyapatite. The activity of the purified enzyme was 2000-3000-fold the casein kinase activity of the cytosol. It had a s20,w of approx. 3 S as determined on sucrose density gradient. After iodination or incubation with [gamma- 32P]ATP, it was analyzed on polyacrylamide gel in the presence of sodium dodecyl sulfate (SDS) and appeared to be composed of a single polypeptide (36 000 +/- 1000 daltons) which self-phosphorylated. In contrast to casein kinase II, casein kinase I preferentially utilized ATP over GTP. The Km value for ATP was determined to be 14 micro M. The Km value for phosvitin was 0.17 mg/ml. Casein kinase I phosphorylated sites different from those of casein kinase II (as shown with ribosomes or SV40 T antigen). Casein kinase I was further characterized by studying its thermal stability. The half-life at 37 degree C was 6 min and 1 min 30 s at 54 degree C. In the presence of two substrates (ATP and phosvitin), the half-life at 54 degree C increased from 1 min 30 s to 4 min. Hemin strongly increased the rate of inactivation of the casein kinase I at 37 degree C in the absence or presence of the substrates. N-ethylmaleimide also inactivated casein kinase I. Phosvitin, though not ATP, protected the enzyme. This observation may indicate that thiols are involved at the binding site of the enzyme for the protein substrate.

Animals↗
PubMed1986

Secretion-coupled protein degradation: studies on mammary casein.

Mammary explants from midpregnant rabbits were cultured for 18 h at 37 degrees C with insulin, prolactin and cortisol. Subsequently, explants were labelled for 2 h with inorganic [32P]phosphate, L-[5-3H]proline or L-[4,5-3H]leucine, washed and chased for up to 3 h. The radiolabelling profile of [32P]casein or [3H]casein during the chase period, obtained by isoelectric focussing or immunoprecipitation indicates extensive destruction of neosynthesized casein. The extent of casein destruction in mammary explants in culture (measured after radiolabelling with L-[5-3H]proline), is inversely related to casein secretion. Least casein degradation is observed in explants after 48 h in culture when casein secretion is maximal (observed histochemically). Subsequently, when the extracellular alveolar lumen is filled with secretion products (72 h), rapid intracellular casein destruction is again observed. When the chase was carried out in the presence of drugs which inhibit degradation and/or secretion, the results indicate that secretion-coupled casein degradation is dependent on an intact functional microfilamentous-microtubular network, casein is not degraded by an autophagosome requiring process, degradation is inhibited by leupeptin, amino-acid analogue containing casein does not undergo secretion-coupled degradation and inhibition of N-glycosylation of intracellular vesicular membrane proteins prevents secretion-coupled degradation. Secretion-coupled protein destruction is discussed in relation to the post-translational regulation of the net production of secretory proteins in eukaryotic cells.

Adenine↗
PubMed1984

Studies of beta-casein content of normal and neoplastic rat mammary tissues by a homologous radioimmunoassay.

We have developed a sensitive homologous radioimmunoassay for rat beta-casein (r-beta-casein) capable of measuring concentrations of beta-casein as low as 0.2 ng/mg of tissue and have measured casein content of rats under several physiological conditions as well as in experimentally induced mammary tumor explants. Changes in content of beta-casein were followed during pregnancy and lactation. Virgin females had 10 ng beta-casein per micrograms of DNA, with a continuous increase in casein content observed throughout pregnancy. At the outset of lactation, beta-casein increased markedly and remained relatively constant throughout lactation. These variations in beta-casein levels indicate that in the rat the synthesis of this protein is under hormonal control and related to the state of differentiation of the mammary gland. The concentration of beta-casein was very low in mammary tumors (0.6 ng/microgram of DNA). Addition of prolactin to organ culture of NMU-induced tumors did not induce significant changes in beta-casein content. These results indicate that NMU-induced tumors have lost at least one of their hormonally regulated functions of cell differentiation, although their hormonal dependence for growth is maintained.

Animals↗
PubMed1990

Anti-adhesive activity of human casein against Streptococcus pneumoniae and Haemophilus influenzae.

The casein fraction of human milk was found to inhibit the attachment of Streptococcus pneumoniae and Haemophilus influenzae human respiratory tract epithelial cells. The inhibitory activity for S. pneumoniae remained after heat and trypsin treatment of the casein and was found in oligosaccharides released from casein. kappa-Casein, which is the most highly glycosylated casein component, inhibited pneumococcal attachment at concentrations similar to the whole casein fraction. The results are consistent with the known recognition of GlcNAc beta 1-3Gal by S. pneumoniae, since human milk and bovine colostrum, which contain GlcNAc, inhibited attachment, but mature bovine milk lacking GlcNAc did not. The effect on H. influenzae was similar to that on S. pneumoniae in that the attachment was inhibited by human casein and bovine colostrum, but not by either mature bovine milk or by the bovine casein fraction. The kappa-casein component of human milk was a less efficient inhibitor of H. influenzae attachment than the whole casein fraction and the free oligosaccharides were inactive. This anti-microbial effect of human casein represents a new mechanism for the protection by breast-milk against respiratory tract infection.

Animals↗
PubMed1999

Casein kinase II-like ectokinase activity on RBL-2H3 cells.

We studied the properties of the ectokinase activity on the outer cell surfaces of RBL-2H3 cells and examined the phosphorylation of exogenous substrates to clarify the substrate specificity of the ectokinases on RBL-2H3 cells. Among the several protein substrates tested, casein was the most strongly phosphorylated with [gamma-32P]ATP, and the net incorporation of 32P into casein was 0.65 pmol P/50 microg/10(6) cells. Casein kinase II peptide was also phosphorylated with [gamma-32P]ATP. The phosphorylation of casein and casein kinase II peptide was almost completely inhibited by the addition of 3 microg/ml of cell-impermeable K252b. Phosphorylation of casein and casein kinase II peptide was also observed by [gamma-32P]GTP. Western blot analysis using anti-casein kinase II antibody revealed a 44-kDa casein kinase band in the membrane fraction and Fc epsilonRI complexes. The immunofluorescence microscopic analysis using anti-casein kinase II antibody showed the existence of casein kinase II on the surface of the cells. This is the first report about the existence of ectokinase on mast cells.

Animals↗
PubMed2001

Primary structure of kappa-casein isolated from mares' milk.

In this work the purification and the complete primary structure of kappa-casein from equine milk are reported for the first time. Mares' milk casein was separated by RP-HPLC into four fractions. Complete primary sequence was obtained by sequence analysis of the protein in the fastest eluting peak isolated by chromatography. This sequence was 95% identical to that reported for the C-terminal portion of the zebras' kappa-casein and showed high similarity with kappa-caseins from sources other than Equidae, confirming that this protein was indeed kappa-casein in equine milk. The presence of post-translational modifications in equine kappa-casein was investigated by mass spectroscopy, after enzymic dephosphorylation. Two main components were found, the smaller component being more abundant. Equine kappa-casein was recognized by a lectin specific for one of the glucosidic bonds in the saccharide moiety of bovine kappa-casein. Sequence comparison with prevision studies showed that the distribution of charged and hydrophobic regions in equine kappa-casein was similar, but not identical, to that found in the bovine protein; these regions are associated with the role of kappa-casein in the formation and stabilization of the micellar structure of casein in milk.

Amino Acid Sequence↗