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At least 919 records · Page 51Linked to original sources
PubMed1997

Protamine-induced hypotension and bradycardia in a cardiac transplant patient.

PURPOSE: The potential for functional reinnervation of the transplanted heart in man is controversial. We report the sudden onset of bradycardia in a cardiac transplant patient following a period of hypotension subsequent to the administration of protamine. Possible mechanisms underlying this response, including reinnervation of the transplanted heart, are assessed. CLINICAL FEATURES: Eight weeks after cardiac transplantation, a patient returned to hospital for a left femoral-tibial artery bypass vein graft. The patient was anaesthetized using general anaesthesia. Upon completion of the procedure, protamine was administered to reverse the heparin-induced anticoagulation. Although administration of a 5.0 mg "test-dose" appeared to be without cardiovascular effect, after an additional 20.0 mg, blood pressure decreased from 98/52 to 62/40 mmHg. After blood pressure reached its nadir, heart rate decreased precipitously from 57 to 29 beats.min-1. CONCLUSIONS: This report demonstrates that heart rate can change considerably in patients who have undergone cardiac transplantation. It is argued that the change in heart rate observed in the present report cannot be explained by reinnervation of the transplanted heart, as the patient had undergone transplantation only eight weeks previously. Rather, we suggest that the change was mediated by mechanisms intrinsic to the transplanted heart and extrinsic to the CNS.

Bradycardia↗
PubMed2003

Protamine mRNA as molecular marker for spermatozoa in semen stains.

Cytological detection of spermatozoa with subsequent DNA analysis is the most important biological evidence in sexual crimes when suitable samples are available. Immunological and enzymatic detection of semen-specific proteins may be helpful but cannot replace specific identification of spermatozoa. We have recently shown that detection of cell-specific gene expression can be used to identify menstrual blood. In this paper we demonstrate that the basic nucleoproteins protamine 1 and 2 are suitable markers for spermatozoa because they are exclusively expressed in the haploid genome and that protamine mRNA can be detected in semen stains by the highly sensitive reverse transcriptase-polymerase chain reaction (RT-PCR). With semi-nested PCR, 10-100 spermatozoa are theoretically sufficient to provide positive amplification results, with hot-start PCR at least 100-1,000 cells are required corresponding to an average semen volume of 0.01-0.1 microl. This new method thus allows specific identification of spermatozoa with molecular biology tools and may broaden the spectrum of investigations in the forensic laboratory.

Adolescent↗
PubMed1987

Pulmonary hypertensive effect of heparin and protamine interaction: evidence for thromboxane B2 release from the lung.

The characteristic pulmonary hypertensive effect of the heparin and protamine interaction has been studied in the isolated pig lung preparation using sequential autologous blood perfusate and dextran perfusate. A significant (p less than 0.001) increase in pulmonary artery pressure at constant flow was seen in 10 of 14 dextran and 12 of 15 blood perfusions. The average increase for dextran was 112 percent and for blood, 109 percent. Antihistamines did not inhibit the response. However, this was abolished in all 11 animals treated with aspirin. In 11 intact swine, thromboxane B2 blood levels increased significantly (p less than 0.01) from 0.46 +/- 0.38 ng/dl to 2.97 +/- 1.5 ng/dl. Thus, pulmonary hypertension associated with protamine reversal of heparinization is associated with prostaglandin release from the lung, and this does not require mediation of platelets or leukocytes.

Animals↗
PubMed1984

Protamine sulfate precipitation: a new assay for the Ah receptor.

The ability of protamine sulfate to effect the quantitative precipitation of 2,3,7,8-[3H]tetrachlorodibenzo-p-dioxin (TCDD):Ah receptor complexes from rat liver cytosol has been developed into a new assay for the identification, quantitation, and characterization of the Ah receptor. The method is reliable, uncomplicated, and rapid, and can be applied to large numbers of samples. The major advantage of the assay is that protamine sulfate appears to selectively precipitate the Ah receptor protein and does not precipitate a number of other proteins that bind [3H]TCDD nonspecifically.

Animals↗
PubMed1990

Reversal of protamine-induced catastrophic pulmonary vasoconstriction by prostaglandin E1.

A case of catastrophic pulmonary vasoconstriction occurring after cardiopulmonary bypass after protamine reversal of heparin treated successfully with intravenous prostaglandin E1 is reported. Systemic hypotension was counteracted by epinephrine given through the left atrium. Protamine-heparin reactions are reviewed and a pathophysiological mechanism for the beneficial effect seen with prostaglandin E1 is proposed.

Alprostadil↗
PubMed1975

Protamine kinase from yeast.

A protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37) which preferentially phosphorylates protamine is purified about 250-fold from the soluble fraction of baker's yeast (Saccharomyces cerevisiae). This enzyme is not sensitive to activation by cyclic nucleotides. Histone is about 5% as active as protamine in the reaction rate. Neither casein, phosvitin nor glycogen phosphorylase is active as substrate. The enzyme is distinguishable from casein kinase of the classical type (Rabinowitz, M. and Lipmann, F. (1960) J. Biol. Chem. 235, 1043-1050) and from adenoshine 3', 5'-monophosphate-dependent protein kinase described earlier (Takai, Y., Yamamura, H. and Nishizuka, Y. (1974) J. Biol. Chem. 249,530-535).

Caseins↗
PubMed1977

Protamine messenger RNA from rainbow trout testis contains the nucleotide sequence A-A-U-A-A-A in an untranslated region.

Full-length, complementary DNAs were prepared to rainbow trout protamine mRNA using reverse transcriptase and were labelled during synthesis by the replacement of dATP by [alpha32P]dATP or dTTP by [alpha32P]dTTP. The 32P-labelled protamine complementary DNAs were digested with T4 endonuclease IV. Fragments from the digests were separated in two dimensions, and those discrete fragments which could be identified from both the A-labelled and T-labelled complementary DNAs were subjected to sequence analysis. The sequences described here all arise from the non-coding region. One pentadecanucleotide contained the sequence A-A-U-A-A-A which has been reported by Proudfoot and Brownlee ((1976) Nature 263, 211-214) to occur in the noncoding regions of six other eukaryotic mRNAs.

Animals↗
PubMed1977

Purification of human immunoglobulin M by affinity chromatography on protamine-Sepharose.

Human IgM has been isolated from plasma by a simple procedure in high yield. The first step was adsorption to protamine-Sepharose and elution by increasing the ionic strength with NaCl. This was followed by two gel filtration steps resulting in a 98% pure IgM in about 30% yield. A somewhat modified procedure could also be used for purification of IgM from Cohn fraction II + III. The purified IgM was found to have a sedimentation constant in agreement with reported values. In immunoelectrophoresis and isoelectric focusing, purified IgM showed the same behaviour as IgM in plasma. Different fragments of IgM were tested for binding to the protamine-Sepharose adsorbent. IgM and Fab were not bound unlike (FC)5, indicating that the sites responsible for binding are located in the Fc part and that several Fc parts are necessary for sufficiently strong binding for adsorption.

Chromatography, Affinity↗
PubMed1979

Improvement in the resolution of human sperm protamines by use of iodoacetamide as alkylating agent.

By use of the neutral alkylating agent iodo [14C1] acetamide instead of ethylene imine or iodoacetate, the resolution of human protamines on gel electrophoresis and ion-exchange chromatography has been improved. Using 20-cm gels, human protamines may be fractionated into seven bands, including the two chromatographically distinct forms of HP1 and a hitherto undetected component HP4. On ion-exchange chromatography, HP2 and the two forms of HP1 may be isolated in sufficient purity for sequence analysis.

Chromatography, Ion Exchange↗
PubMed1979

Protamines and related proteins from spermatozoa of molluscs. Characterization and molecular weight determination by gel electrophoresis.

The spermatozoa of most species of molluscs contain a mixture of proteins with compositions related to those of histones and protamines. The various components present in the spermatozoa of Cryptochiton stellerii, Donax trunculus, Mactra corallina and Gibbula divaricata have been isolated and characterized. The results obtained, taken together with those previously found (Subirana, J.A., Cozcolluela, C., Palau, J. and Unzeta, M. (1973) biochim. Biophys. Acta 317, 364--379), show that in all the molluscs studied one or more arginine-rich components are present. The molecular weight of these proteins varies vary much in different species and is usually much greater than in conventional fish protamines. Conventional histones, as well as lysine-rich proteins of low molecular weight, have also been found in ripe spermatozoa of several species. The molecular weights have been estimated by gel electrophoresis, using polymerized iridine as a standard.

Amino Acids↗
PubMed1992

Protamine kinase phosphorylates eukaryotic protein synthesis initiation factor 4E.

Up to 1 mol of phosphoryl groups was incorporated per mol of eukaryotic protein synthesis initiation factor (eIF) 4E following incubation of purified preparations of this factor with purified preparations of a protamine kinase from bovine kidney cytosol. By contrast, purified preparations of two forms of mitogen-activated protein kinase, casein kinase II and two forms of a distinct autophosphorylation-activated protein kinase exhibited little activity, if any, with eIF-4E. Together with previous observations, the results indicate that the protamine kinase could contribute to the insulin-stimulated phosphorylation of eIF-4E.

Animals↗
PubMed1992

Implications of a non-lamellar lipid phase for the tight junction stability. Part I: Influence of basic amino acids, pH and protamine on the bilayer-hexagonal II phase behaviour of PS-containing PE membranes.

Inverted lipid micelles have been proposed, among other biological functions, to constitute the structural basis of the so-called tight junctions, a special cell cell contact found in epithelia and endothelial, which act as a barrier for the paracellular solute passage. As a model system for the opening and closing of this gate, we investigated the formation of the inverted hexagonal phase (HII phase) in lipid bilayer systems consisting of egg phosphatidylethanolamine (egg PE) and mixed egg PE/bovine brain phosphatidylserine (BBPS) membranes. The formation of the HII phase was modulated by Ca2+ ions, pH, basic amino acids and protamine. The lamellar-HII phase transition temperature TH of pure egg PE membranes at pH 7.0 was lowered with increasing Ca2+ concentration. This effect was attenuated by the presence of 50 mM lysine methyl ester. In the mixed lipid system, this effect was also observed, but even more pronounced. However this effect could be compensated for by raising the Ca2+ concentration from 2 to 10 mM. This was not observed in the pure PE system. In the absence of Ca2+, lysine methyl ester and protamine lowered TH in both monocomponent and mixed lipid systems, whereas lysine caused the opposite effect. The pH-dependence of mixed lipid systems, which were investigated up to a BBPS content of 20 mol%, clearly shows that increasing PS content stabilizes the lamellar phase even at low pH. The results obtained with model membranes are discussed with respect to biological implications of the lamellar-HII phase transition for the modulation of tight junction stability.

Amino Acids↗
PubMed1977

Selective protamine sulphate inactivation of lipoprotein lipase and hepatic lipase in human post-heparin plasma: specific lipase levels in normals and in type I hyperlipoproteinaemia.

The rate of inactivation of triglyceride hydrolase activity by protamine sulphate was determined in pooled, normal, post-heparin plasma. Two distinct first-order rates of inactivation were obtained and the derived constants used to calculate the lipoprotein lipase and hepatic lipase contributions to the total post-heparin triglyceride hydrolase activity in normal controls and in patients with familial hyperchylomicronaemia. The lipoprotein lipase was reduced in the patients whereas the hepatic lipase was normal. There was however a marked age-related increase in the hepatic enzyme activity in normal subjects. Post-heparin lipolytic activity, assayed in vitro against lipoproteins of d less than 1.006 derived from the patients, was markedly reduced in our hyperchylomicronaemic subjects. This assay correlated well with the lipoprotein lipase activity determined by selective protamine sulphate inactivation.

Adolescent↗
PubMed1993

Anion-mediated lysine-arginine interaction. Evidence in Chaetopterus variopedatus sperm protamine.

Chaetopterus ariopedatus sperm protamine is a stable oligomer. Specific amino acid side chain modifications show that the oligomeric structure depends on anion-mediated lysine-arginine interactions. The occurrence of this type of interaction is confirmed by the finding that poly-L-arginine readily forms aggregates with poly-L-lysine or with the native but not with the protamine with carbamylated epsilon-amino groups.

Amino Acids↗
PubMed1983

Thermodynamics and kinetics of co-operative protein-nucleic acid binding. II. Studies on the binding between protamine and calf thymus DNA.

Unspecific binding of a protamine, namely fluorescein-labelled clupeine Z, to double-stranded calf thymus DNA was studied using fluorescence titration methods and chemical relaxation techniques. Both equilibrium and kinetic data have been analysed using general theoretical approaches discussed in the accompanying paper. The results agree well with the predictions made on the basis of a standard co-operative binding model. Basic parameters evaluated are the co-operative binding constant (K), the coefficient measuring co-operative interaction between nearest neighbours (q), the number of nucleotides occupied by one protamine molecule (n) and the rate constant of dissociation at the ends of bound ligand sequences (kD). Values obtained at 20 degrees C, pH 7.5 and 0.4 M-NaCl were K = 5.8 X 10(7) M-1, q = 1700, n = 20 and kD = 0.29 s-1. They have been found to be sensitive to the concentration of added salt (NaCl). This effect apparently reflects the essentially electrostatic nature of the binding process. The results can be satisfactorily described in terms of competitive binding of sodium ions.

Animals↗
PubMed1994

Influence of protamine on the in vitro sensitivity of Pseudomonas aeruginosa to antibiotics.

We have studied the influence of protamine on the sensitivity of three strains of Pseudomonas aeruginosa to various antibiotics: streptomycin, gentamicin, polymyxin B, and beta-lactams. While protamine enhanced the antibacterial action of beta-lactams towards P. aeruginosa, it did not alter the effect of aminoglycosides. The antibacterial power of polymyxin B, on the other hand, was drastically reduced. The observed changes in this strain's sensitivity to antibiotics could be due to changes in the permeability of the outer membrane.

Aminoglycosides↗
PubMed1975

Protamine-precipitated estrogen receptor: a solid-phase ligand exchange assay.

Cytoplasmic estrogen receptor can exist either free (R) or bound to estradiol-17beta (RE). Both forms can be precipitated from cytosols by protamine sulfate. After protamine precipitation R binds 3-H-estradiol-17beta quantitatively at either 0 degrees or 30 degrees, while precipitated RE binds 3-H-estradiol-17beta only at 30 degrees by exchanging with previously bound hormone. Using these observations, we have developed a method for separate determination of both cytoplasmic R and RE. This method should also be applicable for assay of other steroid receptors, especially in cases where interfering components are present in the whole cytosol.

Animals↗
PubMed1987

Isolation and purification of fibrinogen/fibrin degradation products by chromatography on protamine-agarose.

Protamine-agarose chromatography is introduced as a rapid and efficient method for the isolation of fibrinogen/fibrin degradation products from biological fluids such as human plasma. All fibrinogen/fibrin fragments above a MW of 35000, including fragments D and E are quantitatively adsorbed to insolubilized protamine and can be eluted with a buffer containing 0.2 M sodium citrate/citric acid pH 5.3, following previous elution of non-fibrinogen proteins with a buffer containing 0.8 M NaCl at neutral pH. Fragments D and E are separated by stepwise elution. The efficiency of the method is - evaluated by applying it to plasma samples obtained from healthy donors and from patients with clinical and laboratory evidence of disseminated intravascular coagulation.

Chromatography, Agarose↗