Proteins are essential macromolecules that play diverse roles in the physiology and health of horses. They are composed of amino acids and are involved in various biological processes, including tissue growth, repair, and the synthesis of enzymes and hormones. Dietary proteins are a key component of equine nutrition, influencing muscle development, immune function, and overall performance. Horses require a balanced intake of essential amino acids, which must be obtained through their diet, as they cannot be synthesized endogenously. This page compiles peer-reviewed research studies and scholarly articles that explore the types, functions, and dietary requirements of proteins in horses, as well as their impact on equine health and performance.
Hedlund B, Danielson C, Lovrien R.Organic phosphates, ATP, AMP, and 2,3-di-phosphoglycerate (DPG) were interacted with horse oxyhemo-globin. Binding parameters were obtained by means of dialysis equilibrium in buffers at 5°, and from calorimetry at 5 and 25°, all in the pH mom 6.5-7.3. The calorimetric results were evaluated assuming a single strong site, and assuming that the pH shifts which occur in the absence of added salt are due to electrostatic effects and not to changes in side-chain titration, upon mixing samples previously ad-justed to the same pH. There was obtained for ATP and 2,3- diphosphoglycerate. respectivel...
Nishi S, Watabe H, Hirai H.The production of antibody to homologous alpha fetoprotein (AFP) in rabbits, rats, and horses by immunication with human AFP is reported. The antigens were administered subcutaneously 5 times at intervals of 7-10 days. Rabbits and dogs received 1 mg of human AFP/ml of the homologous pooled newborn serum with each injection while the rats received 1/2 of the dose. The horses received 5 mg/ml/injection. 2 weeks after the last injection, antisera were collected and immunologic assays were performed by the Ouchterlony method and the reversed version of the Mancini method. High titered antibodies w...
Thoburn R, Hurvitz AI, Kunkel HG.Various mammalian species contain an anionic serum protein that reacts specifically with native DNA. It is considerably less reactive with single-strand DNA and does not react with monodeoxyribonucleotides, homopolyribonucleotides, or duplexes of homopolyribonucleotides. Synthetic dA.dT was an effective inhibitor of the reaction with native DNA, while Micrococcus luteus DNA and dG.dC were not inhibitory. This protein was encountered in the course of studies on DNA antibodies. Although it reacted with red cells coated with DNA and gave agar precipitation bands, it was clearly distinct from DNA ...
McGuire TC, Crawford TB.Immunoglobulin A (IgA) was demonstrated in equine serum and secretions. This immunoglobulin had a molecular weight extending from 150,000 to 700,000 and reacted with specific antihuman alpha-chain antiserum. Antigenic determinants specific for secretory IgA were demonstrated and found to be absent on serum IgA. Antigen binding activity was detected in IgA from tears. Purified IgA was antigenically distinct from equine IgG, IgM, IgG(T), and aggregating immunoglobulin. Quantitative studies demonstrated that IgA was the predominant immunoglobulin in tears and milk but not in colostrum. The electr...
Gospodarowicz D.A highly purified preparation of PMSG has been obtained from fresh serum and from a commercial preparation. Carbohydrate and amino acid compositions have been determined. The carbohydrate content of PMSG is 46.7% and the molecule is rich in Sialic Acid (13.5%). The apparent molecular weight of PMSG has been determined by SDS polyacrylamide gel electrophoresis. A molecular weight of 53,000 has been found for the unreduced and unalkylated molecule. After reduction and alkylation, the molecular weight fell to 23,000. From these values it has been concluded that PMSG is an oligomeric molecule comp...
Rifkind JM.The reduced glutathione in the erythrocyte was found to inhibit the autoxidation of purified horse hemoglobin. It was observed that much higher concentrations of oxidized glutathione also stabilize hemoglobin. The stabilization by oxidized glutathione most likely involves the formation of a mixed disulfide with the reactive β-93 sulfhydryl groups on the hemoglobin. A similar effect is also observed with N-ethyl- maleimide and HgCl2 which also react with the sulfhydryl groups. The apparent stabilization by reduced glutathione is partially due to the reduction of ferrihemoglobin formed by autox...
Blake CC, Evans PR, Scopes RK.The single peptide chain of 3-phosphoglycerate kinase is folded into two distinct globular units, only one of which seems to be involved in substrate binding.
Norcross NL, Coggins L.The spleens of horses infected with equine infectious anemia contain an antigen that is useful for a diagnostic immunodiffusion test. This antigen was extracted from the spleen by homogenization of the tissue, centrifugation, and precipitation from the supernatant fluid at 50% saturation with (NH(4))(2)SO(4). The antigen was purified by subjecting it to two cycles of electrophoresis in a continuous free-flow electrophoresis cell and finally filtering through a column of Sephadex G-200 gel. The antigen was found to be a small protein with a molecular weight of 27,500 and sedimentation coefficie...
Rakotoarisoa L, Mironneau C, Sayet I, Mironneau J.Specific binding of the Ca++ antagonist desmethoxyverapamil, (-)-[3H]D888, to cell membranes of equine portal vein smooth muscle was inhibited in a concentration-dependent manner by guanosine 5'-O-(gamma-thio)triphosphate and ATP but was little affected by guanosine 5'-O-(beta-thio)diphosphate, noradrenaline or phorbol 12-myristate 13-acetate ester. Inhibition constants for GTP and ATP were in the range of 0.1 to 0.3 mM. From Scatchard plots and dissociation kinetic experiments, it is proposed that D888 high affinity binding sites are transferred into low affinity sites. In intact strips of ra...
Hubbard CD, Shoupe TS.A transient phase for the hydrolysis of indophenyl acetate by the commercial preparation of horse serum cholinesterase was observed on a stopped-flow spectrophotometer. It was found that the transient process is a reaction of the ester with a major component of the preparation and is not caused by the serum cholinesterase enzyme. This noncholinesterase component was isolated and the dependence of its concentration and that of the ester upon the transient liberation of the indophenolate ion were determined. Studies with the isolated component and subsequent analyses have led to the tentative id...
Pain R, Sneddon JC, Cochrane CA.Franz diffusion cells containing uniformly dehydrated equine blood clots to simulate fibrinous eschar were used to measure the rate of debridement of fibrin by novel and traditional wound dressings, under standardised conditions of temperature, pH and humidity. Significant increases in protein breakdown occurred within 24 hours with all the dressings, but not thereafter. In general, dressings hydrated in normal saline were better as debriding agents than dressings hydrated in water. Autolytic debriding agents were 47 per cent more effective than chemical debriding agents; specifically, hydrofi...
Greenaway EC, Sepulveda MF, Cunningham FM, Goode NT.Eosinophils have been implicated in the pathogenesis of the seasonal equine allergic skin disease, sweet itch. Protein kinase C (PKC) is involved in regulating eosinophil function and antigen challenge has been reported to alter PKC isotype expression in blood eosinophils from allergic human subjects. Here we have compared the pattern of PKC isotype expression in eosinophils from sweet itch ponies with that in cells from normal ponies both during the active and inactive phases of the disease. A role for PKC in histamine-induced eosinophil activation was also investigated. Conventional PKCs alp...
Kalpokas I, Martínez MN, Cavestany D, Perdigón F, Mattos RC, Meikle A.We investigated the early effects of the equine embryo on maternal serum concentrations of insulin-like growth factor 1 (IGF1), leptin and adiponectin, uterine immune cells and genes and proteins related to embryo development and the maintenance of pregnancy. Ipsilateral endometrial expression was assessed on Days 7 and 13 after ovulation for the following transcripts: oestrogen receptor ERα (ESR1), progesterone receptor (PGR), progestin and adipoQ receptor family member 5 (PAQR5), oxytocin receptor (OXTR), prostaglandin-endoperoxide synthase 2 (PTGS2), raf-1 proto-oncogene serine/threonine k...
Brems DN, Liu YC, Stellwagen E.The heme iron of horse heart cytochrome c was selectively removed using anhydrous HF. The product, porphyrin c, exhibits the viscosity, far ultraviolet circular dichroic, and fluorescence properties characteristic for native cytochrome c. However, porphyrin c is more susceptible to denaturation by guanidine hydrochloride and by heat than is the parent cytochrome. All of the conformational parameters of porphyrin c exhibit a common reversible transition centered at 0.95 m guanidine hydrochloride at 23 degrees C and pH 7.0. Guanidine denatured porphyrin c refolds in two kinetic phases having tim...
Stratil A, Tomásek V, Bobák P, Glasnák V.Homozygous horse transferrin (Tf O) is highly heterogeneous. In starch gel electrophoresis it gives at least 9 zones. Two main components (2a and 4b) were purified by rivanol and ammonium sulphate precipitation, DEAE-Sephadex chromatography and SP-Sephadex chromatography. Molecular weights of 75 200 and 80 500 for components 2a and 4b, respectively, were determined by sedimentation equilibrium ultracentrifugation. Amino acid compositions of the two components were similar, and there were no differences in the N-terminus (glutamic acid followed by glutamine) and the C-terminus (valine). Differe...
Williams RC.The evolutionary distance between two sets of proteins was estimated using the techniques of Miyata and Yasunaga (1980) and Kimura (1980). Human beta 2-microglobulin was compared with the homologous murine molecule, while human and equine alpha-globin were similarly treated. It was found that a large amount of molecular evolution has occurred in beta 2-microglobulin since its divergence from the common ancestor of mice and humans. Kimura's estimate of evolutionary distance, K, is 0.353, while those of Miyata and Yasunaga are KS = 0.708 and KA = 0.171. The respective values for human and equine...
Aida Y, Okada K, Kageyama R, Amanuma H.Tumor-associated antigens that are expressed in lymphosarcoma B cells of cattle with enzootic bovine leukosis had been analyzed in terms of their reactivity with 13 monoclonal antibodies (MAB). By use of flow cytometry and radioimmunoprecipitation, 1 of the MAB (c143) that recognized a tumor-associated antigen cross-reacted with blood lymphocytes (BL) from various mammalian species. By use of flow cytometry, the c143 MAB reacted with 10 to 49% of BL derived from human beings, mice, dogs, horses, pigs, llamas, sheep, goats, and cattle. Titer of the c143 MAB with BL from horses, pigs, human bein...
Posthuma CC, Pedersen KW, Lu Z, Joosten RG, Roos N, Zevenhoven-Dobbe JC, Snijder EJ.The replication/transcription complex of the arterivirus equine arteritis virus (EAV) is associated with paired membranes and/or double-membrane vesicles (DMVs) that are thought to originate from the endoplasmic reticulum. Previously, coexpression of two putative transmembrane nonstructural proteins (nsp2 and nsp3) was found to suffice to induce these remarkable membrane structures, which are typical of arterivirus infection. Here, site-directed mutagenesis was used to investigate the role of nsp3 in more detail. Liberation of the hydrophobic N terminus of nsp3, which is normally achieved by c...
Kriegshäuser G, Kuechler E, Skern T.Equine rhinitis A virus (ERAV) is a picornavirus which causes an acute respiratory infection in horses worldwide, and virus neutralization (VN) has been the standard method for the detection of ERAV antibody in horse serum. Previous studies have identified recombinant virion protein VP1 (rVP1) purified under native conditions to be of high potential for the development of a diagnostic ERAV enzyme-linked immunosorbent assay (ELISA). This study presents an optimized protocol for the expression and purification of native full-length rVP1. Furthermore, we demonstrate that, upon denaturation, rVP1 ...
Reitnour CM, Salsbury RL.Protein digestion, nitrogen retention, plasma protein, plasma urea, and plasma-free amino acids were determined for ponies fed 3 different protein supplements. Substitution of casein, corn gluten meal, or corn gluten meal plus lysine for a portion of the cornstarch in a low-protein basal ration increased apparent digestion of protein. Substitution with either casein or corn gluten meal plus lysine produced a significant increase (P less than 0.05) in nitrogen retention, whereas the corn gluten meal substitution did not. Nitrogen retention, expressed as percentage of nitrogen absorbed, was incr...
Dai CB, Xiao Y, Lu H, Shen RX, Shao YM.Membrane protein GP90 of China equine infectious anemia virus (EIAV) vaccine strain (DLV) and its parental wild type LN strain were expressed with Bac-to-Bac baculovirus expression system and BALB/c mice were inoculated with purified protein, thereby to explore the availability of protein for differential diagnosis and potential for preparing genetically engineered vaccine. Methods: The authors infected donkey PBMC culture with China EIAV vaccine strain (DLV) and its parental wild type LN strain, extracted its proviral DNA as template, amplified the GP90 of DLV and LN, respectively, and expres...
Miller PA, Lawrence LM.Six mature Quarter Horse mares were used in a crossover design to assess the effect of dietary protein level on metabolic response to exercise. After a 2-wk adaptation period to either a 12.9% (control) or an 18.5% CP (high-protein) diet, each mare performed a standard exercise test. The test consisted of a 15-min exercise period at 4.5 m/sec on a 9% grade motorized treadmill. Exercise resulted in an increase in heart rate (P less than .001), but there was no difference (P less than .05) between dietary treatments. In the jugular vein, lactate increased (P less than .01) from 6.3 to 52.0 mg/10...
Sticht H, Willbold D, Rösch P.Two molecular dynamics (MD) simulations were performed in order to increase the understanding of the dependence of protein conformation on solvent environment. The protein used for these simulations is the transcriptional activator of the equine infectious anemia virus (EIAV-Tat). The structure of this protein has been determined by nuclear magnetic resonance (NMR) in aqueous solution (Willbold et al., Science 264, 1584 (1994)) and in 40% (v/v) trifluoroethanol (TFE) (Sticht et al., Eur. J. Biochem., submitted) showing considerable differences in the stability of the secondary structure elemen...
Hilbert BJ, Rowley G, Antonas KN, McGill CA, Reynoldson JA, Hawkins CD.Sodium hyaluronate was injected into normal horse joints and joints that had undergone an arthrotomy and experimental cartilage damage. The elimination half-life for hyaluronic acid in normal joints was found to be approximately 96 h. The injection caused a non-significant increase (42%) in synovial fluid protein concentration and a fall in the intrinsic viscosity of the fluid. In the arthrotomy group the synovial fluid hyaluronic acid concentration fell after surgery but it was unaffected by the injection of sodium hyaluronate. An initial rise in the intrinsic viscosity of the synovial fluid ...
Kizaki T, Mizuno Y, Takasawa T, Shiokawa H.Analysis of the quantitative precipitin reaction of acylphosphatase from porcine skeletal muscle with rabbit antiserum indicated the presence of at least two antigenic determinants on the porcine enzyme molecule. Immunological cross-reactivities of acylphosphatases from equine and rabbit skeletal muscles were examined. In double immunodiffusion with the antiserum, the precipitin lines of the porcine and equine enzymes completely fused, while the rabbit enzyme gave no precipitin line. The reaction between the 125I-labeled porcine enzyme and its antibody was inhibited to the same extent by the p...
Kolmakova MV, Kuskova ZR, Ratner GM, Laptakova LM.Horse serum albumin has been shown to meet the requirements to protein preparations for microanalysis and thus to be suitable for use in kits of reagents for the radioimmunological determination of insulin and myoglobin, for the determination of tick-borne encephalitis virus antigen by the method of the enzyme immunoassay and for the stabilization of proteins in the hemagglutination test and the hemagglutination inhibition test.
Word TA, Larsen RW.Here the molar volume and enthalpy changes associated with the early events in the folding of ferrocytochrome c (Cc) at high pH have been examined using time resolved photoacoustic calorimetry (PAC). The data reveal an overall volume change of 1.3 ± 0.3 mL mol and an enthalpy change of 13 ± 7 kcal mol occurring subsequent to photodissociation of the unfolded CO bound Cc species in <∼20 ns. Two additional kinetic phases are observed that are associated with non-native His binding (ΔH and ΔV of 2 ± 4 kcal mol and -0.5 mL mol, τ ∼ 2.5 μs ) and Met binding (ΔH an...
Milne EM, Doxey DL, Gilmour JS.The analysis of peritoneal fluid is of value in the differential diagnosis of equine colic but its characteristics have not been evaluated in grass sickness. Peritoneal fluid was collected from 15 normal horses and from 11 cases of medical colic, 11 cases of surgical colic, 20 cases of acute grass sickness and 13 cases of subacute grass sickness. The fluid was analysed for its appearance, total and differential white cell count, specific gravity, total protein concentration and total and intestinal alkaline phosphatase activity. Fluid from cases of medical colic was normal in these respects. S...
Barbey C, Cauchard S, Cauchard J, Laugier C, Hartke A, Petry S.Rhodococcus equi remains a significant pathogen, causing severe pneumonia in foals. The development of vaccines and serologic diagnosis could be greatly facilitated by studying the humoral immune response to this equine pathogen. In this study, a crude extract of R. equi ATCC 33701-secreted proteins combined with the Montanide® ISA70 adjuvant was found to be highly immunogenic in mice with the highest titer of 99,000 on day 42 after the first subcutaneous immunization. This immune response was dependent on the quantity of proteins injected and the presence of adjuvant. By dot-blotting, eight ...
Ueno Y, Uemura R, Niwa H, Higuchi T, Sekiguchi S, Sasaki Y, Sueyoshi M.Equine proliferative enteropathy (EPE) caused by Lawsonia intracellularis is characterized by hypoproteinemia. There are currently no reliable reports that provide a reference value for the total serum protein (TP) concentration to clinically diagnose EPE. The objective of this study was to statistically determine the reference value. Feces and sera of 99 foals with EPE-like clinical signs and of 35 healthy foals were obtained. The samples were used for specific-gene detection of L. intracellularis, TP measurement, and specific-antibody detection against L. intracellularis. Based on these resu...
Dubin A, Potempa J, Turyna B.Horse blood leucocyte cytosol exhibits a broad inhibitory activity against serine proteinases. The purified inhibitor was exposed to investigated enzymes (trypsin, chymotrypsin, elastases and serine proteinase from S. aureus) for variable time and the products were analyzed by gradient polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. The molar ratio I:E, association rate constants k on and inhibition constants Ki for the enzymes and inhibitor were determined. The examined elastases form stable, stoichiometric complexes with the inhibitor (Ki less than 10(-10) M), ...
Thway TM, Wolfe MW.Equids and primates are the only species known to express the placental hormone chorionic gonadotropin (CG). CG is a member of the heterodimeric glycoprotein family and is composed of an alpha subunit linked to a hormone-specific beta subunit. Previously, we have reported that epidermal growth factor (EGF) regulates the equine glycoprotein hormone alpha subunit promoter through a protein kinase C (PKC)/mitogen-activated protein kinase (MAPK) signal transduction pathway in trophoblasts. The current study investigates the regulatory element/factors involved in the induction of equine glycoprotei...
Liu ZQ, Xia J, Wang GL, Kuermanali N.Hyalomma asiaticum tick, an important ectozoic parasite causes tickle, pain, anemia, weight loss, and paralysis in its hosts, which include humans, cattle, sheep, horses, camels, and hares. The 4D8 gene can be a potential vaccine candidate antigen for H. asiaticum. In the present study, we cloned and expressed the 4D8 gene of H. asiaticum from Xinjiang Province. Primers were designed according to the H. asiaticum tick 4D8 gene sequence available in GenBank. The gene was amplified by reverse transcription-polymerase chain reaction and the fragments were subcloned into the prokaryotic expression...
Bewley TA, Li CH.The conformations of human, equine, and porcine pituitary prolactins, as evidenced by various optical properties, have been compared. The alpha-helix contents of all three proteins are essentially identical to each other (60 +/- 5%), as well as to prolactins isolated from other mammalian species. Direct absorption (zero and second-order), difference absorption, fluorescence emission, and circular dichroism spectra suggest that the majority of tyrosine and tryptophan side chains in these three proteins exist in very similar microenvironments within the folded forms of the hormones. Thus, the ge...
Pellegrini A, Hägeli G, Fretz D, von Fellenberg R.An electrophoretic procedure for the qualitative and quantitative assay of protein protease inhibitors is reported. This assay is particularly suited for investigations of crude biological materials when specific antisera are not available. The supporting medium consists of agarose into which denatured fibrinogen is incorporated as the substrate for proteases. The processing then is divided into two steps: (1) electrophoretic resolution of the inhibitor containing material and (2) detection of the inhibitor bands through their protease inhibiting activity. The inhibitor position is thus made v...
Carpenter MA, Broad TE.Transferrin, the iron transport protein of the blood, is highly polymorphic in many species, including the horse. A number of sequence polymorphisms that distinguish several of the variants of horse transferrin are reported here. Previous studies indicated that exons 12 and 15 were likely to be polymorphic. Sequencing regions of exons 12 and 15 from D and R variants revealed 10 nucleotide substitutions that encoded six amino acid replacements. The F1, F2, H2, and * variants were identical to D, and the O variant was almost identical to R, in the regions studied. The data indicated that the hor...
Caffrey CR, Ryan MF.An excretory-secretory (ES) preparation derived from adult Strongylus vulgaris in vitro was assessed for proteolytic activity using azocasein and synthetic, fluorogenic, peptide substrates. Fractionation was by molecular sieve fast protein liquid chromatography (molecular sieve FPLC) and resolution by gelatin-substrate sodium dodecyl sulphate-polyacrylamide gel electrophoresis (gelatin-substrate SDS-PAGE). The cysteine proteinase activator, dithiothreitol (DTT), enhanced azocaseinolysis and hydrolysis of carbobenzoxy-phenylalanyl-arginine-7-amido-4-methylcoumarin (Z-Phe-Arg-NMec) by the ES pre...
Furr M, Chickering WR, Robertson J.To determine normal CSF electrophoresis patterns in horses, and to determine whether the electrophoretic scans from horses with cervical compression differ from those of neurologically normal horses. Methods: 32 horses assigned to 1 of 2 groups: neurologically normal (n = 18) or cervical compression (n = 14). Methods: CSF was collected from 18 neurologically normal horses referred to the Marion duPont Scott Equine Medical Center, and protein electrophoresis was performed to describe the normal equine CSF electrophoretogram. Results of CSF electrophoresis from 14 horses with cervical compressio...
