Detection of one attomole of [Arg8]-vasopressin by novel noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay).
AuthorsHashida S, Tanaka K, Yamamoto N, Uno T, Yamaguchi K, Ishikawa E
JournalJ Biochem (Tokyo)
PubMed ID1778973
'One attomole of [Arg8]-vasopressin (AVP) was detected by a novel noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay). AVP was indirectly biotinylated using N-hydroxysuccinimidobiotin and trapped onto an anti-AVP IgG-coated polystyrene ball. After washing, biotinylated AVP was eluted from the polystyrene ball with HCl and was reacted with 2,4-dinitrophenyl-fluorescein disulfide-bovine ... More
Affinity modulation of platelet integrin alphaIIbbeta3 by beta3-endonexin, a selective binding partner of the beta3 integrin cytoplasmic tail.
AuthorsKashiwagi H, Schwartz MA, Eigenthaler M, Davis KA, Ginsberg MH, Shattil SJ
JournalJ Cell Biol
PubMed ID9182673
'Platelet agonists increase the affinity state of integrin alphaIIbbeta3, a prerequisite for fibrinogen binding and platelet aggregation. This process may be triggered by a regulatory molecule(s) that binds to the integrin cytoplasmic tails, causing a structural change in the receptor. beta3-Endonexin is a novel 111-amino acid protein that binds selectively ... More
Calmodulin and calmodulin-binding proteins in hair bundles.
AuthorsWalker RG, Hudspeth AJ, Gillespie PG
JournalProc Natl Acad Sci U S A
PubMed ID8385344
'Calcium ion plays an important role in the hair cell''s mechanoelectrical transduction process; in particular, Ca2+ controls adaptation to protracted mechanical stimuli. Because calmodulin is a ubiquitous intracellular receptor for Ca2+ and has been shown to accumulate at the tips of stereocilia, we determined its concentration and identified the proteins ... More
Comparative studies of the preparation of immunoliposomes with the use of two bifunctional coupling agents and investigation of in vitro immunoliposome-target cell binding by cytofluorometry and electron microscopy.
'The two coupling agents SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate) and SATA (N-succinimidyl-S-acetylthioacetate) were compared in their efficiency and feasibility to couple monoclonal antibodies (Abs) via thioether linkage to liposomes functionalized by various lipophilic maleimide compounds like N-(3-maleimidopropionyl)-N2-palmitoyl-L-lysine methyl ester (MP-PL), N-(3-maleimidopropionyl)phosphatidylethanolamide (MP-PE), N6-(6-maleimidocaproyl)-N2-palmitoyl-L-lysine methyl ester (EMC-PL), and N-(6-maleimidocaproyl)phosphatidylethanolamine (EMC-PE). The composition of the ... More
Isolation and characterization of thrombin-activated human factor VIII.
AuthorsCurtis JE, Helgerson SL, Parker ET, Lollar P
JournalJ Biol Chem
PubMed ID8119969
'Recombinant human factor VIII (fVIII) was activated by thrombin at pH 7.4, followed by CM-Sepharose chromatography at pH values ranging from 3.5 to 7.4. Optimal coagulant activity was recovered at pH 5.5 and was associated with the isolation of an A1/A2/A3-C1-C2 heterotrimer. The activity was stable at -80 degrees C, ... More
Exosite binding tethers the macromolecular substrate to the prothrombinase complex and directs cleavage at two spatially distinct sites.
AuthorsBoskovic DS, Krishnaswamy S
JournalJ Biol Chem
PubMed ID10984491
'The prothrombinase complex, composed of the proteinase, factor Xa, bound to factor Va on membranes, catalyzes thrombin formation by the specific and ordered proteolysis of prothrombin at Arg(323)-Ile(324), followed by cleavage at Arg(274)-Thr(275). We have used a fluorescent derivative of meizothrombin des fragment 1 (mIIaDeltaF1) as a substrate analog to ... More
Development and testing of radio and enzyme immunoassays for acidic fibroblast growth factor (aFGF).
'Acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor from bovine brain stimulate growth in a variety of tissues in several species. Despite the 55% amino acid sequence homology of the two forms of FGF, a specific immunoassay of aFGF has been developed using a polyclonal antibody raised in ... More
Comparison of four bifunctional reagents for coupling peptides to proteins and the effect of the three moieties on the immunogenicity of the conjugates.
AuthorsPeeters JM, Hazendonk TG, Beuvery EC, Tesser GI
JournalJ Immunol Methods
PubMed ID2499636
'Peptide-carrier conjugates are widely used to raise antipeptide antibodies. In a model system using angiotensin and tetanus toxoid as the peptide and the carrier protein respectively, four cross-linking reagents were employed to study their effect on the immunogenicity of the conjugates. Optimization of the conjugation method for these heterobifunctional reagents, ... More
Liposome targeting to human immunodeficiency virus type 1-infected cells via recombinant soluble CD4 and CD4 immunoadhesin (CD4-IgG).
AuthorsFlasher D, Konopka K, Chamow SM, Dazin P, Ashkenazi A, Pretzer E, Düzgünes N
JournalBiochim Biophys Acta
PubMed ID8075135
'HIV-infected cells producing virions express the viral envelope glycoprotein gp120/gp41 on their surface. We examined whether liposomes coupled to recombinant soluble CD4 (sCD4, the ectodomain of CD4 which binds gp120 with high affinity) could specifically bind to HIV-infected cells. sCD4 was chemically coupled by 2 different methods to liposomes containing ... More
The targeting of immunoliposomes to tumour cells (A431) and the effects of encapsulated methotrexate.
AuthorsJones MN, Hudson MJ
JournalBiochim Biophys Acta
PubMed ID8218324
'Immunoliposomes have been prepared from lipid mixtures of dipalmitoylphosphatidylcholine, wheat germ phosphatidylinositol and a reactive lipid (the m-maleimidobenzoyl-N-hydroxysuccinimide derivative of dipalmitoylphosphatidylethanolamine) which was conjugated to the N-succinimidyl-S-acetylthioacetate (SATA) derivative of a monoclonal antibody (H17E2) raised to human placental alkaline phosphatase (PLAP). The immunoliposomes were prepared by the extrusion technique (VETs) ... More
Cryptomonad algal phycobiliproteins as fluorochromes for extracellular and intracellular antigen detection by flow cytometry.
AuthorsTelford WG, Moss MW, Morseman JP, Allnutt FC
JournalCytometry
PubMed ID11309804
'BACKGROUND: Phycobiliproteins play an important role in fluorescent labeling, particularly for flow cytometry. The spectral properties of R-phycoerythrin (R-PE) and allophycocyanin (APC) have made them the dominant reagents in this class of fluorochromes. In this study, we evaluate a lesser-known but potentially important series of low-molecular weight cryptomonad-derived phycobiliproteins (commercially ... More
A strip liposome immunoassay for aflatoxin B1.
AuthorsHo JA, Wauchope RD
JournalAnal Chem
PubMed ID12033235
'A technique has been developed for the preparation of aflatoxin B1 (AFB1)-tagged liposomes encapsulating a visible dye. These liposomes have several useful potential analytical applications, one of which is demonstrated. A simple plastic-backed nitrocellulose strip is the basis for an assay for detecting AFB1. Samples containing aflatoxin B1 are allowed ... More
A new reagent which may be used to introduce sulfhydryl groups into proteins, and its use in the preparation of conjugates for immunoassay.
AuthorsDuncan RJ, Weston PD, Wrigglesworth R
JournalAnal Biochem
PubMed ID6353995
'A synthesis of the N-hydroxysuccinimide ester of S-acetylthioacetic acid is described. This material is stable when stored dry and has advantages over the currently available reagents used to introduce sulfhydryl groups into a variety of proteins. Proteins modified with this reagent can be used to prepare conjugates for enzyme immunoassay. ... More
A brief survey of methods for preparing protein conjugates with dyes, haptens, and cross-linking reagents.
AuthorsBrinkley M
JournalBioconjug Chem
PubMed ID1616945
Synthesis and purification of horseradish peroxidase-labeled oligonucleotides for tyramide-based fluorescence in situ hybridization.
Authorsvan Gijlswijk RP, van de Corput MP, Bezrookove V, Wiegant J, Tanke HJ, Raap AK
JournalHistochem Cell Biol
PubMed ID10817671
'A method is presented to conjugate horseradish peroxidase (HRP) to oligodeoxynucleotides for fluorescence in situ hybridization assays employing tyramide signal amplification (TSA). HRP is covalently bound to the oligonucleotide by thiol ether linkage and purified by high-performance liquid chromatography. With TSA detection, a single HRP-labeled oligonucleotide probe is sufficient for ... More
Horseradish peroxidase-labeled oligonucleotides and fluorescent tyramides for rapid detection of chromosome-specific repeat sequences.
'We present a sensitive and rapid fluorescence in situ hybridization (FISH) strategy for detecting chromosome-specific repeat sequences. It uses horseradish peroxidase (HRP)-labeled oligonucleotide sequences in combination with fluorescent tyramide-based detection. After in situ hybridization, the HRP conjugated to the oligonucleotide probe is used to deposit fluorescently labeled tyramide molecules at ... More
Binding of factor VIIIa and factor VIII to factor IXa on phospholipid vesicles.
AuthorsDuffy EJ, Parker ET, Mutucumarana VP, Johnson AE, Lollar P
JournalJ Biol Chem
PubMed ID1512239
'The activation of factor X by factor IXa (fIXa) in the presence of phosphatidylcholine-phosphatidylserine (PCPS) vesicles is markedly accelerated by thrombin-activated factor VIII (fVIIIa). The interaction between highly purified fVIIIa and fIXa in this complex was studied fluorometrically at 25 degrees C by using a derivative of D-phenylalanyl-prolyl-arginyl-fIXa which was ... More
Active site-independent recognition of substrates and product by bovine prothrombinase: a fluorescence resonance energy transfer study.
AuthorsBoskovic DS, Troxler T, Krishnaswamy S
JournalJ Biol Chem
PubMed ID14988397
'The conversion of prothrombin to thrombin is catalyzed by prothrombinase, an enzyme complex composed of the serine proteinase factor Xa and a cofactor protein, factor Va, assembled on membranes. Kinetic studies indicate that interactions with extended macromolecular recognition sites (exosites) rather than the active site of prothrombinase are the principal ... More
Mechanisms of delivery of liposome-encapsulated cytosine arabinoside to CV-1 cells in vitro. Fluorescence-microscopic and cytotoxicity studies.
AuthorsBrown PM, Silvius JR
JournalBiochim Biophys Acta
PubMed ID2110480
'Fluorescence microscopy and assays of the cytotoxicity of liposome-encapsulated cytosine arabinoside (araC) have been used to examine the interactions of CV-1 cells with pH-sensitive liposomes, combining phosphatidylethanolamine (PE) with oleic acid or with double-chain protonatable amphiphiles, and with pH-insensitive liposomes combining phosphatidylcholine (PC) and phosphatidylglycerol (PG). Fluorescence-microscopic observations indicate that ... More
Quantitative detection of C-reactive protein using phosphocholine-labelled enzyme or microspheres.
AuthorsDeegan O, Walshe K, Kavanagh K, Doyle S
JournalAnal Biochem
PubMed ID12531203
'C-reactive protein (CRP) is a positive, acute-phase protein. Plasma levels rise dramatically in response to tissue injury or inflammation and fall rapidly after recovery or treatment. Antibody-based human CRP test systems do not readily detect CRP from other animals due to the species specificity of antibodies directed against human CRP. ... More
Protein labeling with fluorescent probes.
AuthorsHolmes KL, Lantz LM
JournalMethods Cell Biol
PubMed ID11060842
Studies of the influence of different cross-linking reagents on the immune response against a B-epitope.
AuthorsDelmas A, Brack A, Trudelle Y
JournalBioconjug Chem
PubMed ID1377494
We have previously shown that the carrier polytuftsin obtained by polycondensation of tuftsin, a naturally occurring macrophage activator, increases significantly the antibody response against a linked B-epitope. In the present work, we have studied the influence of different cross-linking reagents on the quality of the conjugation and on the immune ... More
Fluorochrome-labeled tyramides: use in immunocytochemistry and fluorescence in situ hybridization.
The peroxidase-mediated deposition of hapten- and fluorochrome-labeled tyramides has recently been shown to increase the sensitivity of immunofluorescence and fluorescence in situ hybridization techniques. We have evaluated a number of red, green, and blue fluorescent tyramides for detection of antigens in tissue sections and cytospin preparations and for the detection ... More
Flow cytometric analysis of viability of bull sperm cells.
AuthorsMátyus L, Szabó G, Resli I, Gáspár R, Damjanovich S
JournalActa Biochim Biophys Acad Sci Hung
PubMed ID6545628
Fluorescein diacetate and propidium iodide double fluorescence labeling of animal sperm cells in combination with 55 degrees C treatment for 15 min was used to monitor the viability and heat tolerance of sperm cells by flow cytometry. The applicability of the elaborated test to detect fertility differences has been shown ... More
Liposome immunoassay by long-lived fluorescence detection.
AuthorsOkabayashi Y, Ikeuchi I
JournalAnalyst
PubMed ID9764512
Immunoassay by fluorescence energy transfer from a europium chelate in liposome to allophycocyanin (APC) was demonstrated. Streptavidin or antibody to biotin was bonded to the liposome containing the europium chelate of 2-naphthoyltrifluoroacetone in the bilayer. When the biotin bonded to APC (APC-BT) was added to the prepared liposomes and the ... More
Lectin-mediated targeting of liposomes to a model surface. An ELISA method.
AuthorsHutchinson FJ, Jones MN
JournalFEBS Lett
PubMed ID2455661
Wheat germ agglutinin has been conjugated to the surfaces of sonicated phospholipid liposomes by reacting the protein derivatised with N-succinimidyl-S-acetylthioacetate (SATA) with the m-maleimidobenzoyl-N-hydroxysuccinimide (MBS) derivative of dipalmitoylphosphatidylethanolamine (DPPE) incorporated into the liposomal bilayers. The liposomes as characterised by photon correlation spectroscopy had a weight-average radius of 44 +/- 10 ... More
The characterisation of liposomes with covalently attached proteins.
AuthorsHutchinson FJ, Francis SE, Lyle IG, Jones MN
JournalBiochim Biophys Acta
PubMed ID2914128
The problem of characterising liposomes with covalently attached proteins has been analysed theoretically in terms of a normal weight distribution of liposome diameters. The polydispersity of protein conjugation is considered in terms of the width (standard deviation) of the liposome size distribution. It is shown that the weight-average number of ... More
Relocating the active site of activated protein C eliminates the need for its protein S cofactor. A fluorescence resonance energy transfer study.
AuthorsYegneswaran S, Smirnov MD, Safa O, Esmon NL, Esmon CT, Johnson AE
JournalJ Biol Chem
PubMed ID10026158
The effect of replacing the gamma-carboxyglutamic acid domain of activated protein C (APC) with that of prothrombin on the topography of the membrane-bound enzyme was examined using fluorescence resonance energy transfer. The average distance of closest approach (assuming kappa2 = 2/3) between a fluorescein in the active site of the ... More
Optimized procedures for the coupling of proteins to liposomes.
AuthorsLoughrey HC, Choi LS, Cullis PR, Bally MB
JournalJ Immunol Methods
PubMed ID2391438
A general, optimized method for coupling proteins to liposomes is presented. This procedure utilizes streptavidin covalently coupled to liposomes to allow the subsequent attachment of a variety of biotinated proteins of interest. In the first part of this study, covalent methods for coupling proteins to liposomes which contain the lipid ... More
Magnetic resonance imaging of inducible E-selectin expression in human endothelial cell culture.
AuthorsKang HW, Josephson L, Petrovsky A, Weissleder R, Bogdanov A
JournalBioconjug Chem
PubMed ID11792187
Covalent conjugates of the cross-linked iron oxide nanoparticles (CLIO) and high-affinity (K(d)(app) = 8.5 nM) anti-human E-selectin (CD62E) F(ab')(2) fragments were prepared and tested in vitro to establish feasibility of endothelial proinflammatory marker magnetic resonance (MR) imaging. The conjugates were obtained by using thiol-disulfide exchange reaction between 3-(2-pyridyl)propionyl-CLIO and S-acetylthioacetate-modified ... More
Detection of Cryptosporidium parvum using oligonucleotide-tagged liposomes in a competitive assay format.
AuthorsEsch MB, Baeumner AJ, Durst RA
JournalAnal Chem
PubMed ID11467568
To meet the technical challenge of accurately and rapidly detecting Cryptosporidium parvum oocysts in environmental water, the authors developed a single-use visual-strip assay. The first step in the overall assay procedure involves extracting C. parvum's mRNA coding for heat-shock protein hsp70, followed by amplification using nucleic acid sequence-based amplification (NASBA) ... More
Recent developments in peptide drug delivery to the brain.
AuthorsPardridge WM
JournalPharmacol Toxicol
PubMed ID1523192
Peptide-based therapeutics are highly water-soluble compounds that do not readily enter brain from blood owing to poor transport through the brain capillary endothelial wall, i.e., the blood-brain barrier (BBB). Strategies available for peptide drug delivery to brain include: (a) neurosurgical-based (intraventricular drug infusion, hyperosmotic opening of the BBB); (b) pharmacological-based ... More
A method for selective enrichment and analysis of nitrotyrosine-containing peptides in complex proteome samples.
AuthorsZhang Q, Qian WJ, Knyushko TV, Clauss TR, Purvine SO, Moore RJ, Sacksteder CA, Chin MH, Smith DJ, Camp DG, Bigelow DJ, Smith RD
JournalJ Proteome Res
PubMed ID17497906
Elevated levels of protein tyrosine nitration have been found in various neurodegenerative diseases and age-related pathologies. Until recently, however, the lack of an efficient enrichment method has prevented the analysis of this important low-level protein modification. We have developed a method that specifically enriches nitrotyrosine-containing peptides so that both nitrotyrosine ... More
Protein S alters the active site location of activated protein C above the membrane surface. A fluorescence resonance energy transfer study of topography.
AuthorsYegneswaran S, Wood GM, Esmon CT, Johnson AE
JournalJ Biol Chem
PubMed ID9312108
The location of the active site of membrane-bound activated protein C (APC) relative to the phospholipid surface was determined both in the presence and absence of its cofactor, protein S, using fluorescence resonance energy transfer (FRET). APC was chemically modified to create the FRET donor species, Fl-FPR-APC, with a fluorescein ... More
Active-site-selective labeling of blood coagulation proteinases with fluorescence probes by the use of thioester peptide chloromethyl ketones. I. Specificity of thrombin labeling.
AuthorsBock PE
JournalJ Biol Chem
PubMed ID1634535
In a new strategy for labeling the active sites of serine proteinases with fluorescence probes (Bock, P. E. (1988) Biochemistry 27, 6633-6639), a thioester peptide chloromethyl ketone inhibitor is incorporated into the enzyme active center and used to produce a unique thiol group which provides a site for selective chemical ... More
Active-site-selective labeling of blood coagulation proteinases with fluorescence probes by the use of thioester peptide chloromethyl ketones. II. Properties of thrombin derivatives as reporters of prothrombin fragment 2 binding and specificity of the labeling approach for other proteinases.
AuthorsBock PE
JournalJ Biol Chem
PubMed ID1634536
The behavior of an array of fluorescent human alpha-thrombin derivatives in reporting binding of the fragment 2 domain of prothrombin was characterized as a representative application of the active-site-selective labeling approach to studies of blood coagulation proteinase regulatory interactions. An array of 16 thrombin derivatives was prepared by affinity labeling ... More
The active site of thrombin is altered upon binding to thrombomodulin. Two distinct structural changes are detected by fluorescence, but only one correlates with protein C activation.
AuthorsYe J, Esmon NL, Esmon CT, Johnson AE
JournalJ Biol Chem
PubMed ID1660464
The association of thrombin with thrombomodulin, a non-enzymatic endothelial cell surface receptor, alters the substrate specificity of thrombin. Complex formation converts thrombin from a procoagulant to an anticoagulant enzyme. Structure-function analysis of this change in specificity is facilitated by the availability of two soluble proteolytic derivatives of thrombomodulin, one consisting ... More
The fifth and sixth growth factor-like domains of thrombomodulin bind to the anion-binding exosite of thrombin and alter its specificity.
AuthorsYe J, Liu LW, Esmon CT, Johnson AE
JournalJ Biol Chem
PubMed ID1317850
The domain of thrombomodulin that binds to the anion-binding exosite of thrombin was identified by comparing the binding of fragments of thrombomodulin to thrombin with that of Hirugen, a 12-residue peptide of hirudin that is known to bind to the anion-binding exosite of thrombin. Three soluble fragments of thrombomodulin, containing ... More
Preparation and characterization of conjugates of (modified) human serum albumin and liposomes: drug carriers with an intrinsic anti-HIV activity.
AuthorsKamps JA, Swart PJ, Morselt HW, Pauwels R, De Béthune MP, De Clercq E, Meijer DK, Scherphof GL
JournalBiochim Biophys Acta
PubMed ID8593275
Human serum albumin (HSA) derivatized with cis-aconitic anhydride (Aco-HSA) that was earlier shown to inhibit replication of human immunodeficiency virus type 1 (HIV-1), was covalently coupled to conventional liposomes, consisting of phosphatidylcholine, cholesterol and maleimido-4-(p-phenylbutyryl)phosphatidylethanolamine, using the heterobifunctional reagent N-succinimidyl-S-acetylthioacetate (SATA). The amount of HSA that could be coupled to ... More
The chondroitin sulfate moiety of thrombomodulin binds a second molecule of thrombin.
AuthorsYe J, Esmon CT, Johnson AE
JournalJ Biol Chem
PubMed ID8381406
The role of the chondroitin sulfate moiety of thrombomodulin (TM) in the binding of thrombin to TM has been examined using fluorescent derivatives of thrombin. An anilinonaphthalene-6-sulfonic acid (ANS) dye was attached covalently to the active site histidine of thrombin via a D-Phe-Pro-Arg (FPR) linkage to form ANS-FPR-thrombin. When ANS-FPR-thrombin ... More
Biosensor for dengue virus detection: sensitive, rapid, and serotype specific.
AuthorsBaeumner AJ, Schlesinger NA, Slutzki NS, Romano J, Lee EM, Montagna RA
JournalAnal Chem
PubMed ID11922316
A serotype-specific RNA biosensor was developed for the rapid detection of Dengue virus (serotypes 1-4) in blood samples. After RNA amplification, the biosensor allows the rapid detection of Dengue virus RNA in only 15 min. In addition, the biosensor is portable, inexpensive, and very easy to use, making it an ... More
The active site of factor IXa is located far above the membrane surface and its conformation is altered upon association with factor VIIIa. A fluorescence study.
AuthorsMutucumarana VP, Duffy EJ, Lollar P, Johnson AE
JournalJ Biol Chem
PubMed ID1512240
The topography of membrane-bound blood coagulation factor IXa (fIXa) and the nature of its interaction with its cofactor, factor VIIIa (fVIIIa), were examined using fluorescent derivatives of fIXa. A fluorescein dye was covalently attached to the active-site histidine of fIXa via a D-Phe-Pro-Arg tripeptide tether to form Fl-A-FPR-fIXa; similarly, a ... More
The location of the active site of blood coagulation factor VIIa above the membrane surface and its reorientation upon association with tissue factor. A fluorescence energy transfer study.
AuthorsMcCallum CD, Hapak RC, Neuenschwander PF, Morrissey JH, Johnson AE
JournalJ Biol Chem
PubMed ID8910432
The topography of membrane-bound blood coagulation factor VIIa (fVIIa) was examined by positioning a fluorescein dye in the active site of fVIIa via a tripeptide tether to yield fluorescein-D-phenylalanyl-L-prolyl-L-arginyl-fVIIa (Fl-FPR-fVIIa). The location of the active-site probe relative to the membrane surface was determined, both in the presence and absence of ... More
Active site selective labeling of serine proteases with spectroscopic probes using thioester peptide chloromethyl ketones: demonstration of thrombin labeling using N alpha-[(acetylthio)acetyl]-D-Phe-Pro-Arg-CH2Cl.
AuthorsBock PE
JournalBiochemistry
PubMed ID3219359
The feasibility of a new approach to incorporation of spectroscopic probes into the active sites of certain serine proteases has been demonstrated. The method is based on inactivation of a serine protease with a thioester derivative of a peptide chloromethyl ketone. The thiol group generated by reaction of the covalent ... More
Development of a procedure for coupling the homing device glu-plasminogen to liposomes.
AuthorsHeeremans JL, Kraaijenga JJ, Los P, Kluft C, Crommelin DJ
JournalBiochim Biophys Acta
PubMed ID1384709
The aim of this study was to find a suitable way of coupling the homing-device glu-plasminogen to the outside of liposomes. The described procedure is based on the reaction of thiol-groups introduced in the protein with thiol-reactive groups of the liposome. Details on the thiolation of proteins with the reagent ... More