A nondestructive method for peptide bond conjugation of antigenic haptens to a diphtheria toxoid carrier, exemplified by two antisera specific to acetolactate synthase.
AuthorsMarcussen J, Poulsen C
JournalAnal Biochem
PubMed ID1799217
A method for the preparation of an activated protein carrier is described: Protein carboxyl groups are transformed into N-hydroxysulfosuccinimide esters, a structure that will react with primary amino groups under amide bond formation. Although the activated ester is unstable under aqueous conditions, a significant amount of hapten molecules can be ... More
Zero-length crosslinking between subunits delta and I of the H(+)-translocating ATPase of chloroplasts.
AuthorsBeckers G, Berzborn RJ, Strotmann H
JournalBiochim Biophys Acta
PubMed ID1385976
'Treatment of spinach thylakoids with 1-ethyl-3-(dimethylaminopropyl)-carbodiimide (EDC)/N-hydroxysulfosuccinimide (sulfo-NHS) induced formation of a zero-length crosslink of an apparent molecular mass of 38 kDa. This product was shown, by immunodetection, to consist of subunit delta of CF1 and subunit I of CF0. The crosslink was isolated by preparative SDS gel electrophoresis and ... More
Targeting glucose oxidase at aspartate and glutamate residues with organic two-electron redox mediators.
AuthorsBattaglini F, Koutroumanis M, English AM, Mikkelsen SR
JournalBioconjug Chem
PubMed ID7849073
'The bimolecular rate constants for the reactions of five organic two-electron redox mediators with reduced glucose oxidase (GOx) were determined by measuring voltammetric electrocatalytic currents at glassy carbon electrodes in the presence of excess glucose under anaerobic conditions. The mediators studied were thionine, brilliant cresyl blue, azure A, daunomycin, and ... More
Modulation of the immune response to pneumococcal type 14 capsular polysaccharide-protein conjugates by the adjuvant Quil A depends on the properties of the conjugates.
AuthorsVerheul AF, Versteeg AA, De Reuver MJ, Jansze M, Snippe H
JournalInfect Immun
PubMed ID2925240
'Streptococcus pneumoniae type 14 capsular polysaccharide-bovine serum albumin (S14PS-BSA) conjugates were prepared by water-soluble-carbodiimide-mediated condensation with or without the use of N-hydroxy-sulfosuccinimide. The immunogenicities of the capsular polysaccharide (S14PS) and of the conjugates were studied in (CBA/N x BALB/c)F1 mice and in female BALB/c mice. The response in these mice ... More
Fast electron transfer from cytochrome c6 and plastocyanin to photosystem I of Chlamydomonas reinhardtii requires PsaF.
AuthorsHippler M, Drepper F, Farah J, Rochaix JD
JournalBiochemistry
PubMed ID9174349
'To study the function of the PsaF subunit of photosystem I (PSI), the interactions between plastocyanin or cytochrome c6 and PSI isolated from wild-type and a PsaF-deficient mutant of Chlamydomonas reinhardtii have been examined using cross-linking techniques and flash absorption spectroscopy. We show that efficient electron transfer from both plastocyanin ... More
Basic carboxyl groups of hemoglobin S: influence of oxy-deoxy conformation on the chemical reactivity of Glu-43(beta).
AuthorsRao MJ, Acharya AS
JournalJ Protein Chem
PubMed ID1675854
'The gamma-carboxyl groups of Glu-43(beta) and Glu-22(beta) of hemoglobin-S (HbS), two intermolecular contact residues of deoxy protein, are activated by carbodiimide at pH 6.0. The selectivity of the modification by the two nucleophiles, glycine ethyl ester (GEE) and glucosamine, is distinct. Influence of N-hydroxysulfosuccinimide, a reagent that rescues carbodiimide-activated carboxyl ... More
Carbodiimide modification enhances activity of pig pancreatic phospholipase A2.
AuthorsFerreira JP, Sasisekharan R, Louie O, Langer R
JournalEur J Biochem
PubMed ID8055932
'Pig phospholipase A2, pig iso-phospholipase A2 and bovine pancreatic phospholipase A2 were reacted in solution with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, in the presence of N-hydroxysulfosuccinimide, at pH 7. The influence of micellar protectants was analyzed. In the presence of n-hexadecylphosphocholine, the losses of activity in micellar diheptanoyl-lecithin were 80, 35, and 10% in ... More
Amide cross-linking: an alternative to glutaraldehyde fixation.
AuthorsGirardot JM, Girardot MN
JournalJ Heart Valve Dis
PubMed ID8894992
'BACKGROUND AND AIMS OF THE STUDY: A new fixation method for bioprosthetic tissues is being developed, which does not utilize the standard glutaraldehyde treatment. This method, referred to as Ultifix, uses a coupler and a coupling enhancer with or without one or more coupling agents. It fixes the tissue by ... More
Ni(II) and Ni(I) forms of pentaalkylamide derivatives of cofactor F430 of Methanobacterium thermoautotrophicum.
AuthorsHamilton CL, Ma L, Renner MW, Scott RA
JournalBiochim Biophys Acta
PubMed ID1648401
'A series of pentaalkylamide forms of F430 and of its 12,13-diepimer have been generated and characterized. Carbodiimide-assisted N-hydroxysulfosuccinimide activation of all five peripheral carboxylates of the F430 macrocycle allows nucleophilic attack by a number of primary amines (RNH2, R- = CH3-, CH3CH2-, CF3CH2-, CH3(CH2)3-) generating the pentaalkylamide derivatives. The identity ... More
Succinylated polylysine as a possible link between an antibody molecule and deferoxamine.
'Modification of antibodies with chelating polymers may be helpful for radioimmunoimaging, radioimmunotherapy, and NMR tomography. Succinylated polylysine was activated with carbodiimide/N-hydroxysulfosuccinimide in dimethyl sulfoxide and isolated as a dry solid. Sulfosuccinimide-esterified polymer was used for the two-stage coupling of an amino-containing chelating agent (deferoxamine) to monoclonal R11D10 (IgG) or its ... More
Protein-heparin interactions measured by BIAcore 2000 are affected by the method of heparin immobilization.
AuthorsOsmond RI, Kett WC, Skett SE, Coombe DR
JournalAnal Biochem
PubMed ID12423639
'Surface plasmon resonance (SPR) biosensors such as the BIAcore 2000 are a useful tool for the analysis of protein-heparin interactions. Generally, biotinylated heparin is captured on a streptavidin-coated surface to create heparinized surfaces for subsequent binding analyses. In this study we investigated three commonly used techniques for the biotinylation of ... More
Enhancement by N-hydroxysulfosuccinimide of water-soluble carbodiimide-mediated coupling reactions.
AuthorsStaros JV, Wright RW, Swingle DM
JournalAnal Biochem
PubMed ID3740412
Water-soluble carbodiimides are frequently employed in coupling or conjugation reactions, e.g., to link a peptide immunogen to a carrier protein. However, their utility is limited by low coupling yields obtained under some conditions. We have found that addition of N-hydroxysulfosuccinimide to such reactions can greatly enhance the yields obtained. ... More
Shape effects of nanoparticles conjugated with cell-penetrating peptides (HIV Tat PTD) on CHO cell uptake.
AuthorsZhang K, Fang H, Chen Z, Taylor JS, Wooley KL,
JournalBioconjug Chem
PubMed ID18690739
In order to probe the nanoparticle shape/size effect on cellular uptake, a spherical and two cylindrical nanoparticles, whose lengths were distinctively varied, were constructed by the selective cross-linking of amphiphilic block copolymer micelles. Herein, we demonstrate that, when the nanoparticles were functionalized with the protein transduction domain of human immunodeficiency ... More
N-hydroxysulfosuccinimide active esters: bis(N-hydroxysulfosuccinimide) esters of two dicarboxylic acids are hydrophilic, membrane-impermeant, protein cross-linkers.
AuthorsStaros JV
JournalBiochemistry
PubMed ID7126526
We have synthesized and characterized N-hydroxysulfosuccinimide, a new hydrophilic ligand for the preparation of active esters. We have incorporated this ligand into two new protein cross-linking reagents, 3,3'-dithiobis(sulfosuccinimidyl propionate) and bis(sulfosuccinimidyl) suberate. In experiments with rabbit muscle aldolase, it is demonstrated that both of these reagents are highly efficient protein ... More
Comparison between different strategies of covalent attachment of DNA to glass surfaces to build DNA microarrays.
DNA microarray is a powerful tool allowing simultaneous detection of many different target molecules present in a sample. The efficiency of the array depends mainly on the sequence of the capture probes and the way they are attached to the support. The coupling procedure must be quick, covalent, and reproducible ... More
A novel FRET approach for in situ investigation of cellulase-cellulose interaction.
AuthorsWang L, Wang Y, Ragauskas AJ,
JournalAnal Bioanal Chem
PubMed ID20694721
A novel real-time in situ detection method for the investigation of cellulase-cellulose interactions based on fluorescence resonance energy transfer (FRET) has been developed. FRET has been widely used in biological and biophysical fields for studies related to proteins, nucleic acids, and small biological molecules. Here, we report the efficient labeling ... More
Highly efficient immunoliposomes prepared with a method which is compatible with various lipid compositions.
AuthorsHolmberg E, Maruyama K, Litzinger DC, Wright S, Davis M, Kabalka GW, Kennel SJ, Huang L
JournalBiochem Biophys Res Commun
PubMed ID2610692
Monoclonal antibody was conjugated to N-glutaryl-phosphatidylethanolamine in the presence of octylglucoside by using N-hydroxysulfosuccinimide as a carboxyl-activation reagent. The conjugated antibody was then incorporated into liposomes by a simple dialysis method. The method is mild and is compatible with various lipid compositions of the liposomes. We have prepared immunoliposomes containing ... More
Mapping of the protein import machinery in the mitochondrial outer membrane by crosslinking of translocation intermediates.
AuthorsSöllner T, Rassow J, Wiedmann M, Schlossmann J, Keil P, Neupert W, Pfanner N
JournalNature
PubMed ID1530986
Mitochondria contain a complex machinery for the import of nuclear-encoded proteins. Receptor proteins exposed on the outer membrane surface are required for the specific binding of precursor proteins to mitochondria, either by binding of cytosolic signal recognition factors or by direct recognition of the precursor polypeptides. Subsequently, the precursors are ... More
Stabilization of human prostatic acid phosphatase by coupling with chondroitin sulfate.
AuthorsLuchter-Wasylewska E, Dulinska J, Ostrowski WS, Torchilin VP, Trubetskoy VS
JournalBiotechnol Appl Biochem
PubMed ID1905136
Human prostatic acid phosphatase (PAP) (EC 3.1.3.2) was covalently linked to chondroitin sulfate A from whale cartilage. In order to bind the protein amino groups with the preactivated carboxyl groups of chondroitin sulfate, 1-ethyl-3-(3'-dimethylaminepropyl)carbodiimide and N-hydroxysulfosuccinimide were used as coupling agents. The product was soluble and enzymatically active. The activity ... More
Identification of the amino acids involved in the functional interaction between photosystem I and ferredoxin from Synechocystis sp. PCC 6803 by chemical cross-linking.
AuthorsLelong C, Sétif P, Lagoutte B, Bottin H
JournalJ Biol Chem
PubMed ID8144501
Ferredoxin isolated from the cyanobacterium Synechocystis sp. PCC 6803 has been chemically cross-linked to purified photosystem I from the same organism. The reaction was catalyzed by N-ethyl-3-(3-dimethylaminopropyl)carbodiimide in the presence of N-hydroxysulfosuccinimide. A short reaction time and neutral pH values can be used in the presence of the two reagents, ... More
Novel carboranyl amino acids and peptides: reagents for antibody modification and subsequent neutron-capture studies.
AuthorsVaradarajan A, Hawthorne MF
JournalBioconjug Chem
PubMed ID1772906
A new alpha-amino acid derivative incorporating the 1,2-dicarba-closo- dodecarborane(12) cage, namely 5-(2-methyl-1,2-dicarba-closo-dodecarborane(12)-1-yl)- 2-aminopentanoic acid (2), was synthesized by the alkylation of the benzophenone Schiff's base of glycine methyl ester with 3-(2-methyl-1,2-dicarba-closo-dodecaborane(12)-1-yl)pr opyl iodide (8). This amino acid was employed in the synthesis of peptide derivatives such as 19-21 using solid-phase ... More
Reactions of N-hydroxysulfosuccinimide active esters.
AuthorsAnjaneyulu PS, Staros JV
JournalInt J Pept Protein Res
PubMed ID3667072
N-Hydroxysulfosuccinimide esters are reactive functional groups employed in a variety of protein modification reagents, especially cross-linking reagents. For these compounds, hydrolysis is the most important reaction competing for reaction of the esters with nucleophilic groups in proteins. We have employed model compounds to investigate the rates of hydrolysis of N-hydroxysulfosuccinimide ... More
Interaction of specific platelet membrane proteins with collagen: evidence from chemical cross-linking.
AuthorsKotite NJ, Staros JV, Cunningham LW
JournalBiochemistry
PubMed ID6466633
Two recently developed membrane-impermeant cross-linkers, 3,3'-dithiobis(sulfosuccinimidyl propionate) (DTSSP) and bis(sulfosuccinimidyl) suberate (BS3), have been used to examine the interaction of human platelets with collagen. Reaction of human platelets with either of the two cross-linking reagents at micromolar concentrations completely inhibited platelet aggregation in response to collagen but not in response ... More
Direct measurements of multiple adhesive alignments and unbinding trajectories between cadherin extracellular domains.
AuthorsSivasankar S, Gumbiner B, Leckband D
JournalBiophys J
PubMed ID11259289
Direct measurements of the interactions between antiparallel, oriented monolayers of the complete extracellular region of C-cadherin demonstrate that, rather than binding in a single unique orientation, the cadherins adhere in three distinct alignments. The strongest adhesion is observed when the opposing extracellular fragments are completely interdigitated. A second adhesive alignment ... More
Epidermal growth factor (EGF)-like repeats of human tenascin-C as ligands for EGF receptor.
AuthorsSwindle CS, Tran KT, Johnson TD, Banerjee P, Mayes AM, Griffith L, Wells A
JournalJ Cell Biol
PubMed ID11470832
Signaling through growth factor receptors controls such diverse cell functions as proliferation, migration, and differentiation. A critical question has been how the activation of these receptors is regulated. Most, if not all, of the known ligands for these receptors are soluble factors. However, as matrix components are highly tissue-specific and ... More
A facile, water-soluble method for modification of proteins with DOTA. Use of elevated temperature and optimized pH to achieve high specific activity and high chelate stability in radiolabeled immunoconjugates.
AuthorsLewis MR, Raubitschek A, Shively JE
JournalBioconjug Chem
PubMed ID7873659
We have developed a method for attachment of the macrocyclic chelating agent 1,4,7,10-tetraazacyclododecane N,N',N",N"'-tetraacetic acid (DOTA) to proteins by activation of a single carboxyl group with N-hydroxysulfosuccinimide (sulfo-NHS). The sulfo-NHS active ester of DOTA was prepared in a single step using 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC), and DOTA conjugates of cytochrome c and ... More
The interaction of monomeric actin with two binding sites on Acanthamoeba actobindin.
AuthorsBubb MR, Lewis MS, Korn ED
JournalJ Biol Chem
PubMed ID1995634
Actobindin was previously shown to be an 88-residue polypeptide (Mr 9761) with an internal tandem repeat of 33-34 amino acids. Sedimentation equilibrium experiments have confirmed this Mr for native actobindin. Pyreneglyoxal-labeled actobindin had a similar Mr by sedimentation equilibrium analysis and bound to actin in a manner qualitatively similar to ... More
Voltammetric DNA biosensor for cystic fibrosis based on a modified carbon paste electrode.
AuthorsMillan KM, Saraullo A, Mikkelsen SR
JournalAnal Chem
PubMed ID7978297
Carbon paste electrodes modified by the inclusion of either octadecylamine or stearic acid were used as solid phases to which DNA was covalently bound. Immobilized DNA was detected by voltammetry of solutions containing submillimolar quantities of Co(bpy)3(ClO4)3, Co(phen)3(ClO4)3, and Os(bpy)3-Cl2 (bpy = 2,2'-bipyridine; phen = 1,10-phenanthroline), all of which associate ... More
Chemo-mechanical leak formation in human erythrocytes upon exposure to a water-soluble carbodiimide followed by very mild shear stress. II. Chemical modifications involved.
AuthorsThelen P, Deuticke B
JournalBiochim Biophys Acta
PubMed ID3179292
The chemical reactions underlying the chemo-mechanical leak formation in human erythrocytes upon treatment with the carboxyl-modifying reagent 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and subsequent minimal shearing, described in the preceding paper (Thelen, P. and Deuticke, B. (1988) Biochim. Biophys. Acta 944, 285-296), are here characterized in more detail. The capacity to form leaks ... More
Sequence-selective biosensor for DNA based on electroactive hybridization indicators.
AuthorsMillan KM, Mikkelsen SR
JournalAnal Chem
PubMed ID8238927
Deoxyribonucleic acid was covalently immobilized onto oxidized glassy carbon electrode surfaces that had been activated using 1-[3-(dimethylamino)-propyl]-3-ethylcarbodimide hydrochloride and N-hydroxysulfosuccinimide. This reaction is selective for immobilization through deoxyguanosine (dG) residues. Immobilized DNA was detected voltammetrically, using tris (2,2'-bipyridyl)cobalt(III) perchlorate and tris (1,10-phenanthroline)cobalt(III) perchlorate (Co(bpy)3(3+) and Co(phen)3(3+). These complexes are reversibly ... More