Phylogenetic variations in the calcium-dependent electrophoretic shift of alpha-lactalbumin.

alpha-Lactalbumin undergoes a calcium-dependent electrophoretic shift at pH 8.3. When Ca2+ is removed by a chelator, the mobility of the protein increases, reflecting the exposure of negative electrical charges. The shift, however, is not observed by electrophoresis in the presence of SDS, which demonstrates that alpha-lactalbumin does not undergo a measurable conformational change upon debinding of Ca2+. Relative electrophoretic mobilities vary from 1.0 (no shift) to 1.4 among alpha-lactalbumins of different orders of mammals. The differences suggest a variable number of gram atoms of Ca2+ bound to alpha-lactalbumin or substitution of amino acid Ca2+ ligands in the calcium-binding loop.

Differences between individuals in high-molecular weight glycoproteins from mammary epithelia of several species.

Milk fat globules are secreted by envelopment in plasma membrane of the lactating cell. SDS-gel electrophoresis of proteins from this membrane has revealed differences between milk donors in two mucin-like glycoproteins. One of these glycoproteins resolves in 3% acrylamide stacking gel and the other in 4% running gel. The proteins vary in number of bands (one or two) and band mobilities. This polymorphism arises, at least in part, from expression of hypervariable genes. In this study, gel electrophoretic evidence of similar polymorphism in glycoproteins from cow, chimpanzee, horse and human milks is presented. In distinction to the other species, the cow expressed only one of these proteins which was detected in the running gel at Mr 180,000 to 200,000. The electrophoresis pattern for this protein from six cows was highly varied with respect to number (one or two) and position of bands. Peanut agglutinin, wheat germ agglutinin and concanavalin A all were bound specifically by bands of the bovine glycoprotein. Binding of concanavalin A distinguishes the bovine protein from the two human glycoproteins. Further studies of species differences should help shed light on the evolution of these unique glycoproteins and their possible functions in mother and young.

The calcium-dependent electrophoretic shift of alpha-lactalbumin, the modifier protein of galactosyl transferase.

alpha-Lactalbumin, the modifier protein of galactosyl transferase in the synthesis of lactose by the mammary gland, has been shown to undergo a Ca2+-dependent electrophoretic shift. Such shifts, characteristic of most calcium modulated proteins, are related to gross conformational changes upon binding calcium when detected in the presence of detergent (SDS-PAGE). However, we detected the calcium shift for alpha-lactalbumin using non-denaturing PAGE (ND-PAGE) where electrical charge changes are observed upon binding calcium. In order for a shift to be observed between the apo and calcium bound protein, calcium ion binding to proteins must have minimal dissociation constants (Kdiss) of 10(-7) M; alpha-lactalbumin is reported to bind calcium at Kdiss = 10(-10) to 10(-12) M. The electrophoretic shift identifies alpha-lactalbumin in complex milk whey patterns of many species of mammals.

Interspecies variation in milk composition among horses, zebras and asses (Perissodactyla: Equidae).

Milk samples of four species of wild equids (onager, Equus hemionus onager; mountain zebra, E. zebra hartmannae; plains zebra, E. burchelli; Przewalski horse, E. caballus przewalskii) and two domesticated equids (ass, E. asinus; pony, E. caballus) were analysed. At mid to late lactation the milks of all species were very similar, containing on average 10-12% total solids, 1-2% fat, 1.6-1.8% true protein, 6-7% 'lactose', 0.3-0.5% ash, 0.08-0.12% calcium, 0.04-0.07% phosphorus and a calculated energy content of 2.0-2.4 kJ/g. Milk samples collected in the first 2 weeks after birth showed elevated levels of total solids and protein, and some had reduced 'lactose' levels, but there were no observable trends in milk composition during mid to late lactation (1-12 months post partum). It was concluded that these closely related species produce milks that are nearly identical in gross composition and that the domestic horse is a representative model for the study of equid lactation.

Casein and lactose concentrations in milk of 31 species are negatively correlated.

Casein and lactose concentrations in milks of various species vary by at least 1-2 orders of magnitude, yet no single species secretes both components at either the high or the low end of the ranges. This pattern of variation could result from evolutionary adaptations in a single secretory mechanism.

Ascorbic acid content of neotropical plant parts available to wild monkeys and bats.

The ascorbic acid content of foliage available to wild primates and bats in Panama (in transition between wet and dry seasons) was lower than that of temperate zone foliage but higher than that of most fruits and vegetables. Intakes of ascorbic acid (mg/kg b.wt/day) by wild primates and frugivorous bats in Panama are much greater than that of most human populations.

Lactational performance of various mammalian species.

Lactational performance varies widely among mammalian species. Lactation plays a variable role in the overall reproductive strategy. The fraction of total growth and development of the young contributed by lactation is minimal in such species as guinea pigs but is a very high proportion in some marsupials and primates. Although the overall daily capacity for transfer of energy for many species is rather constant at about 125 kcal/kg body weight.70, the proportions of nutritive constituents in milk differ greatly among species and vary during lactation.

Interrelationships of constituents and partition of salts in milk samples from eight species.

Skim-milk samples from rat, rabbit, pig, sheep, goat, horse and man were analysed for lactose, casein and the total and ultrafilterable concentrations of the main salts. Results are compared with data for the cow. The ultrafiltrate concentrations of Ca and Mg were positively correlated with that of citrate and the colloidal concentrations of Ca, Mg and citrate were positively correlated with that of Pi, suggesting that common, general, principles determine the partition of salts in milks. Casein concentration in the skim-milks was inversely related to that of lactose in accordance with a recent theoretical treatment of the principal mechanism of milk secretion. It is postulated that much of the interspecific diversity in milk composition can be explained by adaptations in a single secretory mechanism.

Ontogenetic development of L-gulonolactone oxidase activity in several vertebrates.

Activity of L-gulonolactone oxidase (EC 1.1.3.8) in livers of fetal Rattus norvegicus and Mus musculus was detectable on the 18th day of gestation, increased rapidly to maxima at 15 and 5 days postpartum for the two species, respectively, and thereafter declined to adult levels. L-Gulonolactone oxidase was not detectable in liver or kidney of fetal guinea pigs at any stage of development. Near-term fetal snowshoe hares had higher activities of liver L-gulonolactone oxidase than observed in a large sample of adults. L-Gulonolactone oxidase was detectable in chicken (Gallus gallus) embryos by the sixth day of incubation, increased rapidly in the kidney with no discontinuity at hatching, reached a maximum at about the 35th day from the beginning of incubation, and then declined to adult levels. Barn swallow (Hirundo rustica) embryos appeared to synthesize little if any L-ascorbic acid; nestlings had considerably higher levels of L-gulonolactone oxidase than adults. Tadpoles of three species of frogs had appreciable levels of L-gulonolactone oxidase activity.

Ascorbic acid turnover and excretion in two arboreal marsupials and in laboratory rabbits.

Rates of turnover and routes of excretion of 1-14C-ascorbic acid were compared among laboratory rabbits and two species of arboreal marsupials. Ringtail possums (Pseudocheirus peregrinus) exhibited higher blood ascorbate levels than either brushtail possums (Trichosurus vulpecula) or rabbits, as well as greater pool sizes, shorter half-times and higher total entry rates of ascorbic acid. These differences were due partly to the high ascorbic acid content of Eucalyptus foliage, the ringtail's diet; when the dietary contribution of ascorbate was taken into account, endogenous entry rates (i.e. 'synthesis') were not significantly different among species. The main excretory route of 14C from 1-14C-ascorbate was expired CO2 in all three species. Estimates to total pool size of ascorbate by isotope dilution were higher than those by tissue analysis. This was considered to be due to excretion of 14C during equilibration of injected 1-14C-ascorbate with the body pool.

Estimation of free myo-inositol in milks of various species and its source in milk of rats (Rattus norvegicus).

Free myo-inositol content of milk of rats and other rodents fed standard laboratory chow is 80 mg/100 g as compared to about 4 mg/100 g for cow milk. Feeding experiments with graded amounts of myo-inositol demonstrated that the myo-inositol content of milk is strongly influenced by dietary intake. Radioactivity from either [carbon-14] glucose or [hydrogen-3] myo-inositol injected intraperitoneally was incorporated rapidly into milk myo-inositol. Thus, both mammary biosynthesis and active transport contribute to the high myo-inositol in rat milk, but the latter is quantitatively the more important.

Isolation and characterization of beta- and gamma-caseins from horse milk.

Three groups of casein components were isolated from horse milk. Group I is almost insoluble at acid and neutral pH, and is rather heterogeneous on alkaline gels with or without sodium dodecyl sulphate. Group II shows strong similarity to beta-casein from other species, as concluded from its amino acid composition and its N- and C-terminal sequences. This group consists of five electrophoretically distinguishable forms, all containing ester phosphate groups but no carbohydrate. Group III is composed of C-terminal fragments of the beta-like (group II) fraction and probably arises from the action of a plasmin-like enzyme present in horse milk. It does not contain phosphate or carbohydrate. Homology of this group with bovine gamma-caseins is demonstrated. Both beta- and gamma-like caseins are more soluble at 4 degrees C than at room temperature.

Stereospecificity of L-myo-inositol-1-phosphate synthase for nicotinamide adenine dinucleotide.

Partially purified preparations of L-myo-inositol.-1-phosphate synthase (EC 5.5.1.4) from testis and mammary gland of laboratory rats (Rattus norvegicus) were used to show that this enzyme is specific for the pro-S hydrogen at C-4 of its cofactor, nicotinamide adenine dinucleotide (NAD). pro-S specificity of the first step (reversible oxidation of glucose 6-phosphate to 5-ketoglucose 6-phosphate) was proved by showing that tritium is transferred from [pro-S-4-3H]NADH but not from [pro-R-4-3H]NADH to glucose 6-phosphate when they are incubated with enzyme. That the stereospecificity in the second oxidation--reduction step (reduction of myo-inosose-2 1-phosphate to myo-inositol 1-phosphate) is the same as in the first step was shown by demonstrating that tritium from [5-3H]glucose 6-phosphate is incorporated into myo-inositol but not into NAD+ and that tritium from [4-3H]NAD+ is not incorporated into myo-inositol

Effect of chymosin action on the hydrodynamic diameter of casein micelles.

Quasi-elastic light scattering shows an initial decrease of about 5 nm in the hydrodynamic radius of casein micelles after adding chymosin, assuming the decrease to be equal for all micelles. This is consistent with the hypothesis that casein micelles have a hairy outer layer that is partly made up of the caseino-macropeptide part of kappa casein.

Effect of ascorbic acid deficiency on the in vivo synthesis of carnitine.

The effects of ascorbate deficiency on carnitine biosynthesis was investigated in young male guinea pigs. Liver and kidney carnitine levels were not affected by the deficiency, but scorbutic animals had 50% less carnitine in heart and skeletal muscle than control animals. Labeled carnitine precursors, 6-N-tri-methyl-L-lysine and 4-N-trimethylaminobutyrate, both of which require ascorbate for their enzymatic hydroxylation, were injected into the vena cava of control, pair-fed and scorbutic animals. The distribution of isotope in compounds present in the liver and kidney after 1 h was determined. The uptake of trimethyllysine by the liver was less than 2% in 1 h, while the kidney took up approx. 20% of the 14C. Control and pair-fed animals converted trimethyllysine to kidney trimethylaminobutyrate 8--10 times as well as did scorbutic animals. Trimethylaminobutyrate hydroxylase, present in the liver but almost absent from the kidney, converted nearly all of substrate taken up by the liver to carnitine in both the scorbutic and control animals.

Ascorbic acid biosynthesis in the mammalian kidney.

The egg-laying mammals (Prototheria) synthesize L-ascorbic acid only in kidney, as is characteristic of reptiles. Bandicoots (Marsupialia) synthesize it in both kidney and liver. 2 other species of marsupials (kangaroos) synthesize it primarily in liver, but some individuals also synthesize in kidney.

The composition of human milk.

Mature human milk contains 3%--5% fat, 0.8%--0.9% protein, 6.9%--7.2% carbohydrate calculated as lactose, and 0.2% mineral constituents expressed as ash. Its energy content is 60--75 kcal/100 ml. Protein content is markedly higher and carbohydrate content lower in colostrum than in mature milk. Fat content does not vary consistently during lactation but exhibits large diurnal variations and increases during the course of each nursing. Race, age, parity, or diet do not greatly affect milk composition and there is no consistent compositional difference between milks from the two breasts unless one is infected. The principal proteins of human milk are a casein homologous to bovine beta-casein, alpha-lactalbumin, lactoferrin, immunoglobulin IgA, lysozyme, and serum albumin. Many enzymes and several "minor" proteins also occur. The essential amino acid pattern of human milk closely resembles that found to be optimal for human infants. Possible special functions of milk proteins and enzymes other than as a source of amino acids, are as yet largely speculative. The principal sugar of human milk is lactose but 30 or more oligosaccharides, all containing terminal Gal-(beta 1,4)-Glc and ranging from 3--14 saccharide units per molecule are also present. These may amount in the aggregate to as much as 1 g/100 ml in mature milk and 2.5 g/100 ml in colostrum. Some of them may function to control intestinal flora because of their ability to promote growth of certain strains of lactobacilli. Human milk fat is characterized by high contents of palmitic and oleic acids. the former heavily concentrated in the 2-position and the latter in the 1- and 3-positions of the triglycerides. Fatty acid composition of milk fat varies somewhat with the composition of diet, particularly the fatty acids which it supplies. Phospholipids, amounting in the aggregate to about 75 mg/100 ml, include phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl serine, phosphatidyl inositol, and sphingomyelin. The principal mineral constituents of human milk are Na, K, Ca, Mg, P, and Cl. Calcium concentrations reported in various studies vary from 25--35 mg/100 ml. Phosphorus at 13--16 mg/100 ml is much more constant but is lower in proportion to casein and calcium than in milks of most other species. Iron, copper, and zinc contents of human milk vary considerably. A long list of other trace elements has been reported. About 25% of the total nitrogen of human milk represents nonprotein compounds including urea, uric acid, creatine, creatinine, and a large number of amino acids. Of the latter, glutamic acid and taurine are prominent. All of the vitamins, except K, are found in human milk in nutritionally significant concentrations.

PIP: A complete and authentic picture of the qualitative and quantitative composition of the milk of Homo sapiens is presented. Older original references are reexamined along with data prublished during the last 2 decades. Mature human milk is made up of 3%-5% fat, 0.8%-0.0% protein, 6.9%-7.2% carbohydrate calculated as lactose, and 0.2% mineral constituents expressed as ash. The energy content is 60-75 kcal/100ml. Protein content is considerably higher and carbohydrate content lower in colostrum than in mature milk. Fat content does not vary consistently during lactation but exhibits large diurnal variations and increases during the course of each nursing. Race, age, parity, or diet fail to have a great affect on milk composition. There is no consistent compositional difference between milks from the 2 breasts unless 1 breast is infected. The principal proteins of human milk are a casein homologous to bovine B-casein, a-lactalbumin, lactoferrin, immunoglobulin IgA, lysozyme, and serum albumin. Lactose is the principal sugar of human milk. Human milk fat is characterized by high contents of palmitic and oleic acids, the former heavily concentrated in the 2-position and the latter in the 1- and 3-positions of the triglycerides. The principal mineral constituents of human milk are Na, K, Ca, Mg, P, and C1. About 25% of the total nitrogen of human milk represents nonprotein compounds. These include urea, uric acid, creatine, creatinine, and a large number of amino acids.

Comparative aspects of milk proteins.

The current status of knowledge of the major proteins of milks of various species is evaluated. Most of the non-bovine milk proteins are homologous with the recognized families of those of Bos taurus, alpha S1-caseins, alpha S2-caseins, beta-caseins, kappa-caseins, beta-lactoglobulins, and alpha-lactalbumins, each family representing a separate genetic locus specific to the mammary gland. No prominent milk protein not homologous to one of these families has yet been discovered in milk of any species. Genetic polymorphism resulting from substitutions in the polypeptide chains and various degrees of post-translational phosphorylation, glycosylation, and proteolysis have been identified in milk proteins of several species. Total protein production ranges among species from about 0.5 to 10 g/d per kg0.75 maternal weight. Proportions of the several proteins vary greatly among species, but few accurate analytical data are available except for total casein and total whey protein contents.