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PubMed1996

[Comparative analysis of RFLP of mitochondrial DNA in Eastern Slavic populations in Russia].

Data on restriction fragment length polymorphism (for entire mtDNA molecules--BamHI, DraI, EcoRI, HindIII, PstI, PvuII, ScaI; for the 16,081-16,545 bp segment of mtDNA--AvaII, BamHI, EcoRV, HaeIII, HinfI, KpnI, RsaI, Sau3AI) in eastern Slavs of Magadan is presented. By Sau3AI/BclI digestion analysis, a new polymorphic variant of mtDNA in Caucasoid populations with the C-T nucleotide substitution in position 16147 bp was revealed. The absence of significant differences in mitotypic diversity of Slavic samples from Magadan (h = 0.921 in Russians, h = 0.898 in Ukrainians) was observed. By comparative analysis of mtDNA RFLP, the presence of genetic differences between eastern Slavic populations of Magadan, Tomsk, and the cities of the European region of Russia was demonstrated.

DNA, Mitochondrial↗
PubMed1997

Mechanisms of C5a and C3a complement fragment-induced [Ca2+]i signaling in mouse microglia.

Microglial cells are activated in response to brain insults; the mechanisms of this process are not yet understood. One of the important signaling mechanisms that might be involved in microglia activation is related to changes in the intracellular calcium concentration ([Ca2+]i). Using fluo-3 microfluorimetry, we have found that external application of the complement fragment C5a (4-10 nM) induced [Ca2+]i elevation in microglial cells in situ in corpus callosum slices. Similarly, application of complement fragments C5a (0.1-10.0 nM) or C3a (100 nM) generates biphasic [Ca2+]i transients composed of an initial peak followed by a plateau in cultured microglia. Incubation of microglial cells for 30 min with pertussis toxin (PTX; 1 microgram/ml) inhibited both C5a- and C3a-triggered [Ca2+]i responses, suggesting the involvement of PTX-sensitive G-proteins in the signal transduction chain. Removal of Ca2+ ions from the extracellular solution eliminated the plateau phase and limited the response to the initial peak. The restoration of the extracellular Ca2+ concentration within 30-60 sec after the beginning of the complement fragment-induced [Ca2+]i elevation led to the recovery of the plateau phase. Inhibition of the endoplasmic reticulum Ca2+ pumps with 500 nM thapsigargin transiently increased the [Ca2+]i and blocked the [Ca2+]i signals in response to subsequent complement fragment application. Our data suggest that complement factors induce [Ca2+]i responses by Ca2+ release from internal pools and subsequent activation of Ca2+ entry controlled by the filling state of the intracellular Ca2+ depots.

Animals↗
PubMed1997

Analysis of the in vitro secondary structure of cucumber mosaic virus satellite RNA.

The solution conformation of two variants of cucumber mosaic virus satellite RNA (CMV satRNA) was analyzed using several enzymatic and chemical probes. Ribonuclease T1 and nuclease S1 were used to map unpaired nucleotides, and nuclease V1 was used to detect double-stranded, or stacked, bases. Chemical probing with dimethylsulphate and diethylpyrocarbonate also identified unpaired and unstacked nucleotides, respectively. Modified or cleaved positions were identified by direct gel electrophoresis of radioactively labeled RNA, or by analysis of DNA sequence patterns generated by primer-extension with reverse transcriptase. Additional information was obtained by a gel-fractionation method under nondenaturing conditions for the identification of base paired fragments. On these data, a model for the in vitro secondary structure of CMV satRNA is proposed. Results support the existence of a complex structure with 51% of nucleotides involved in base pairs (40 G:C, 28 G:U, and 18 A:U pairs). Several structural elements, numbered I-VI, were defined, and interactions between separate domains are suggested. Comparisons of experimental data and a formerly reported secondary structure model for CMV satRNA support the validity of the structure we propose.

Base Composition↗
PubMed2005

Chemically-assisted fragmentation combined with multi-dimensional liquid chromatography and matrix-assisted laser desorption/ionization post source decay, matrix-assisted laser desorption/ionization tandem time-of flight or matrix-assisted laser desorption/ionization tandem mass spectrometry for improved sequencing of tryptic peptides.

Derivatization of tryptic peptides using an Ettan CAF matrix-assisted laser desorption/ionization (MALDI) sequencing kit in combination with MALDI-post source decay (PSD) is a fast, accurate and convenient way to obtain de novo or confirmative peptide sequencing data. CAF (chemically assisted fragmentation) is based on solid-phase derivatization using a new class of water stable sulfonation agents, which strongly improves PSD analysis and simplifies the interpretation of acquired spectra. The derivatization is performed on solid supports, ZipTip(microC18, limiting the maximum peptide amount to 5 microg. By performing the derivatization in solution enabled the labeling of tryptic peptides derived from 100 microg of protein. To increase the number of peptides that could be sequenced, derivatized peptides were purified using multidimensional liquid chromatography (MDLC) prior to MALDI sequencing. Following the first dimension strong cation exchange (SCX) chromatography step, modified peptides were separated using reversed-phase chromatography (RPC). During the SCX clean up step, positively charged peptides are retained on the column while properly CAF-derivatized peptides (uncharged) are not. A moderately complex tryptic digest, prepared from six different proteins of equimolar amounts, was CAF-derivatized and purified by MDLC. Fractions from the second dimension nano RPC step were automatically sampled and on-line dispensed to MALDI sample plates and analyzed using MALDI mass spectrometry fragmentation techniques. All proteins in the derivatized protein mixture digest were readily identified using MALDI-PSD or MALDI tandem mass spectrometry (MS/MS). More than 40 peptides were unambiguously sequenced, representing a seven-fold increase in the number of sequenced peptides in comparison to when the CAF-derivatized protein mix digest was analyzed directly (no MDLC-separation) using MALDI-PSD. In conclusion, MDLC purification of CAF-derivatized peptides significantly increases the success rate for de novo and confirmative sequencing using various MALDI fragmentation techniques. This new approach is not only applicable to single protein digests but also to more complex digests and could, thus, be an alternative to electrospray ionization MS/MS for peptide sequencing.

Amino Acid Sequence↗
PubMed1996

A morphometric study of the epithelium lining the rat intrahepatic biliary tree.

BACKGROUND/AIMS: Morphological and functional heterogeneity of intrahepatic bile duct epithelial cells has been suggested in situ and in isolated cholangiocytes. The aim of this study was to evaluate if: (a) bile ducts, when isolated, maintain morphometric parameters similar to ducts in situ, (b) cellular organelles show heterogeneity in ducts of different size, and (c) some features permit different classes of bile ducts to be distinguished. METHODS: Studies in situ were conducted on normal liver processed for light or electron microscopy. Data were also obtained from preparations of intrahepatic biliary tree isolated from rat liver. The whole biliary tree was cut at different levels to obtain bile ducts of different diameter. The diameter of ducts, the number of lining cells, the size and the area of individual cells, the nucleo/cytoplasmic ratio, the volume density of mitochondria, endoplasmic reticulum, Golgi complex and lysosomes have been evaluated. RESULTS: The diameter of intrahepatic bile ducts ranged from 5 to 100 micrograms and the area of lining cells ranged from 8 to 100 micrograms2. A highly significant linear relationship existed between duct diameter and bile duct epithelial cell area (r = 0.97, p < 0.001) or number of lining cells (r = 0.96, p < 0.001). The volume density of mitochondria ranged from 7.58 +/- 2.0% of cytoplasmic volume in the smallest isolated bile ducts to 8.50 +/- 2.7% in the largest (p = NS). The volume density of lysosomes was low and was not significantly different in ducts of different size. Rough endoplasmic reticulum was inconspicuous in the smallest ducts and increased only slightly in the largest. The inverse relationship between the nucleo/cytoplasmic ratio and duct diameter was striking (r = -0.78, p < 0.001). All morphometric data were equivalent if bile ducts were evaluated in situ or in isolated fragments. Taken together, the data allowed bile ducts to be classified into 3 classes: < 10, 10-50, and > 50 micrograms in diameter. DISCUSSION: Our data show that (a) isolated bile ducts maintain morphometric characteristics similar to the tissue in situ, (b) a low grade of morphological heterogeneity is evident for intracellular organelles in ducts of different diameter and (c) the diameter of ducts, the number of lining cells and especially the nucleo/cytoplasmic ratio may indicate the origin of fragments examined where functional studies are being considered.

Animals↗
PubMed2001

Mining the mammalian genome for artiodactyl systematics.

A total of 7,806 nucleotide positions derived from one mitochondrial and eight nuclear DNA segments were used to provide a robust phylogeny for members of the order Artiodactyla. Twenty-four artiodactyl and two cetacean species were included, and the horse (order Perissodactyla) was used as the outgroup. Limited rate heterogeneity was observed among the nuclear genes. The partition homogeneity tests indicated no conflicting signal among the nuclear genes fragments, so the sequence data were analyzed together and as separate loci. Analyses based on the individual nuclear DNA fragments and on 34 unique indels all produced phylogenies largely congruent with the topology from the combined data set. In sharp contrast to the nuclear DNA data, the mtDNA cytochrome b sequence data showed high levels of homoplasy, failed to produce a robust phylogeny, and were remarkably sensitive to taxon sampling. The nuclear DNA data clearly support the paraphyletic nature of the Artiodactyla. Additionally, the family Suidae is diphyletic, and the nonruminating pigs and peccaries (Suiformes) were the most basal cetartiodactyl group. The morphologically derived Ruminantia was always monophyletic; within this group, all taxa with paired bony structures on their skulls clustered together. The nuclear DNA data suggest that the Antilocaprinae account for a unique evolutionary lineage, the Cervidae and Bovidae are sister taxa, and the Giraffidae are more primitive.

Animals↗
PubMedDate not supplied

[Parasite communities and a fish spawn].

High values of dominance index, low values of evenness and Shannon index are characteristic of component parasite communities of prespawning (Prosopium cylindraceum) and spawn migratory fishes (Coregonus autumnalis, Oncorhynchus nerka). Autogenic specialists are dominant in the component parasite communities of C. autumnalis. The component parasite communities of P. cylindraceum and O. nerka are dominant in a content of generalist species. The O. nerka parasite communities are communities "sentenced to death". Low values of dominance index, high values of evenness abd Shannon index are peculiar to component parasite communities of prespawning and spawning cyprinid fishes (Phoxinus phoxinus, Oreoleuciscus humilis). Autogenic specialists are dominant. Increase of dominance index and decrease of two other indexes characterize the postspawning period. The same tendency characterises infracommunities. Thus, the component parasite communities respond differently to the fish spawn of the cyprinid and coregonid/salmonid fishes. Similarity is in the dominance of autogenic specialists in one case only (C. autumnalis). These differences are defined by the mode of fish stock formation for spawn. The cyprinids congregate for spawning and move apart for foraging and vice versa is observed in the coregonids and salmonids. Autogenic specialists dominance seems to be the important adaptation to reduce a negative effect to host during spawning. Bush and Kennedy in 1994 established that "parasites live in patches (host individuals) and fragments (host populations)". They consider the host fragmentation as "hedging your bets against extinction" due to frequency and magnitude anthropogenic factors, which increase fragmenting of host populations. This conclusion was made for a species level. From the other hand the fragmentation is a natural feature for a single host population too. Such fragmentation is a host population structure. The role of fragmentation is obvious from the data on component parasite communities during fish spawn. The fragmentation decreases in the cyprinids and increases in the salmonids that leads to the growing of the cyprinid component parasite communities diversity and to declining the salmonid component parasite communities diversity. Nevertheless the role of host population structure in a component parsite community structure is not obvious, because parasites are able or not able to "recognise" different host subpopulation groups. It is well known from data on parasite species population biology. Such recognizable subpopulation groups or groups can be a "real fragment" for the parasite community. The question is what parameters could be used for this purpose on a component community level. Host population age structure can be used as an example, because the age groups are one of the invariable population characters. Value of Shannon index for component parasite communities of spawn migratory O. nerka (5+) is similar to that of fishes of 1+ age. Difference is statistically insignificant. It is insignificant between the parasite communities of 2+ and 3+ age groups too. Fishes of these two groups could be defined as a real united fragment. It spite of similarity between the fishes of 5+ age group and fish of 1+ age group they are not united fragment. The parasite community of 1+ age fishes is not stabilized yet and one of 5+ age is a community "sentenced to death". Thus the structuring of O. nerka freshwater parasite communities are defined by 3 real host age fragments: 1+ age group, 2 and 3+ age group, 4+ age group. It looks as that Shannon index is suitable parameter to study a parasite communities structure.

Age Factors↗
PubMed2002

Discordance between the effect of modulators of calcium on staurosporine-induced apoptosis and staurosporine-induced actin degradation.

Staurosporine produced DNA fragmentation characteristic of apoptosis and a dramatic alteration of cell structure that include alterations of cytoskeletal actin and cytoplasmic condensation with vacuolization. These changes were not induced by chelerythrine, a more specific PKC inhibitor than staurosporine. The calcium chelator, BAPTA, significantly reduced staurosporine-induced DNA fragmentation but did not affect staurosporine-induced changes in cytoskeletal actin. These data suggest that DNA fragmentation and actin degradation in apoptosis, induced by staurosporine, are under different regulatory control by calcium.

Actins↗
PubMed2005

Identification of proteolytic fragments from ErbB-2 that induce apoptosis.

The receptor tyrosine kinase ErbB-2 plays an important role in cell proliferation and differentiation as well as oncogenesis. We have found that ErbB-2 kinase domain fragmentation is important for the induction of apoptosis. Exogenous expression of peptides derived from the ErbB-2 kinase domain induces cells death with the hallmarks of apoptosis. In contrast, transfection of the ErbB-2 carboxy-terminal domain did not induce apoptosis. We have identified a 37-residue segment from the ErbB-2 kinase N-terminal lobe that can strongly induce apoptosis in transfected cells. Cell death was not blocked by the pan-caspase inhibitor z-VAD-FMK. Similar fragments derived from several other receptor tyrosine kinases also induce cell death. These data imply that proteolytic fragmentation of tyrosine kinases liberates apoptotic fragments that can accelerate cell death.

Animals↗
PubMed1983

Fragmentation of inhaled 238PuO2 particles in lung.

Beagle dogs were exposed by inhalation to monodisperse aerosols of 238PuO2. Autoradiographs of lung sections were studied for nine dogs sacrificed from 4 to 730 days after exposure. Fragmentation of particles in lung was observed autoradiographically. A simple mathematical model fitted to the fragmentation and lung retention data for dogs suggests that lung clearance from intact particles (65%) and particle fragments (32%) were important to the removal of 238PuO2 deposited in the lung by inhalation. The increased surface area resulting from fragmentation should increase dissolution and risk to other organs from irradiation by translocated plutonium.

Aerosols↗
PubMedDate not supplied

Identification and mass spectrometric sequence studies of fragments of l-asparaginase produced during freeze/thaw cycling.

L-Asparaginase isolated from Er. chrysanthemi was found to lose activity upon exposure to consecutive freeze/thaw cycles. The cause(s) for this loss of activity were investigated using multiple techniques. SEC using UV, RI and light scattering detectors and SDS-PAGE indicated that the l-asparaginase molecule fragments upon exposure to repeated freezing and thawing cycles. Following up on this information, mass spectrometry was used to identify the fragments as small peptides of molecular weight 615 Da, 1424 Da and 1665 Da. Automated Edman sequencing of the frozen and thawed mixture confirmed the presence of fragments and contributed some sequence information. Mass spectral data and sequence studies of these fragments in conjunction with the known sequence of the molecule placed all the fragments within the last 28 C-terminal amino acids. A study of this region using the published 3 dimensional x-ray crystallographic structure of l-asparaginase revealed that the C-terminal region is exposed and can interact with water. The IBI MacVector program "Protein Tool Box" predicted that this region is hydrophilic, has a high surface probability and a strong tendency to interact with water. Both tendencies suggest a potential for bond stress during freeze/thaw cycling. This region is not involved at the catalytic core of the enzyme, but fragmentation in this area may result in unfolding and denaturation of the monomer followed by subsequent aggregation into large, insoluble entities and the loss of enzymatic activity.

Amino Acid Sequence↗
PubMed1995

Temporal profile of in situ DNA fragmentation after transient middle cerebral artery occlusion in the rat.

We measured the temporal profile and anatomic distribution of cells exhibiting DNA fragmentation at various durations of reperfusion after middle cerebral artery (MCA) occlusion in the rat. Focal cerebral ischemia was induced in male Wistar rats (n = 62) using an intraluminal monofilament blockade of the MCA. After 2 h of MCA occlusion, the animals were killed at different durations of reperfusion (0.5, 3, 6, 9, and 12 h and 1, 2, 4, 7, 14, 21, and 28 days, n = 4 per time point). Sham-operated rats (n = 4) and normal rats not subjected to any surgical procedure (n = 4) were used as controls. Coronal brain sections (5 microns) were analyzed, using an in situ ApopTag kit, hematoxylin and eosin, and immunohistochemical double-staining methods. Six rats subjected to 2 h of MCA occlusion were killed at 24 h for measurement of DNA fragmentation by gel electrophoresis. Our data indicate that within a coronal section, DNA fragmentation was present in zero to three cells in each hemisphere of normal and sham-operated rats as well as in the contralateral hemisphere of ischemic rats. The number of cells exhibiting DNA fragmentation increased as early as 0.5 h (8 +/- 6), peaked at 24-48 h (213 +/- 59), and persisted for 4 weeks (10 +/- 2) after onset of reperfusion (p < 0.01). Groups of cells exhibiting DNA fragmentation (> 95% neurons) were located primarily in the inner boundary zone of the infarct. With use of gel electrophoresis, purified DNA obtained from the ischemic tissue exhibited the characteristic nucleosome ladder pattern associated with apoptosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2006

Hsp70 chaperone machine remodels protein aggregates at the initial step of Hsp70-Hsp100-dependent disaggregation.

Exposure to temperatures over a certain limit leads to massive protein aggregation in the cell. Disaggregation of such aggregates is largely dependent on the Hsp100 and Hsp70 chaperones. The exact role of the Hsp70 chaperone machine (composed of DnaK, DnaJ, and GrpE) in the Hsp100-dependent process remains unknown. In this study we focused on the Hsp70 role at the initial step of the disaggregation process. Two different aggregated model substrates, green fluorescent protein (GFP) and firefly luciferase, were incubated with the Hsp70 machine resulting in efficient fragmentation of large aggregates into smaller ones. Our data suggest that the observed fragmentation is achieved first by extraction of polypeptides from aggregates in Hsp70 chaperone machine-dependent manner and not by direct fragmentation of large aggregates. In the absence of Hsp100 (ClpB) these "extracted" polypeptides were not able to fold properly and promptly reassociated into new aggregates. The extracted GFP molecules were efficiently recognized and sequestered by a molecular trap, the mutant GroEL D87K, which binds stably to unfolded but not to native polypeptides. The binding of extracted GFP molecules to the GroEL trap prevented their reaggregation. We propose that the Hsp70 machine disentangles polypeptides from protein aggregates prior to Hsp100 action.

Endopeptidase Clp↗
PubMed2002

In vivo phosphorylation of Drosophila melanogaster nuclear lamins during both interphase and mitosis.

To study phosphorylation of D. melanogaster nuclear lamins in vivo, we used Kc tissue culture cells. Kc cells contain products of both lamin genes, the lamin Dm0 gene encoding constitutive polypeptides expressed in almost all cell types and the developmentally regulated lamin C gene. We grew Kc cells in low phosphate medium and labelled them with (32P(H3PO4. To obtain mitotic cells we used vinblastine to arrest cells in metaphase. Cells were collected, washed, lysed and resultant extracts fractionated in the presence of protein phosphatase inhibitors. D. melanogaster proteins were then denatured by boiling in SDS plus DTT, followed by immunoaffinity chromatography and SDS-PAGE purification. As anticipated, we found that a CNBr fragment derived from the N-terminal part of lamin Dm0-derivatives (amino acid residues 2-158; fragment A) was phosphorylated during both interphase and mitosis. Interphase but not mitotic phosphorylation was found on an internal CNBr fragment (derived from the end of the central rod domain and the first part of the C-terminal lamin tail; amino acid residues 385-548; fragment D). Interphase only phosphorylation was also detected on another CNBr fragment derived from the extreme C-terminal portion of lamin Dm0-derivatives (amino acid residues 549-622; fragment E). To supplement these data, we used 2-D tryptic peptide mapping followed by phosphorImager analysis. We routinely detected at least seven 'spots' derived from interphase lamins but only a single mitotic lamin phosphopeptide.

Amino Acid Sequence↗
PubMed1979

Narcolepsy: a family study.

We obtained medical and psychological assessments and 48-hr polysomnographic recordings on five sisters, three of whom had narcolepsy. Of the three, two were identical twins. All three narcoleptic sisters cited emotional stress and environmental demands for sustained performance as the major factors which aggravated their symptoms, and corresponding to this, the illness followed a different life course in each of the three. Most striking were the differences between the twin sisters in clinical symptoms and polysomnographic signs. One sister suffered from all the symptoms of narcolepsy and her sleep recording showed the typical sleep onset REM periods of the disease. Her twin suffered only from excessive daytime drowsiness and her sleep recording was normal--at least by the usual criteria. The sleep of all three narcoleptic sisters, however, was significantly more fragmented than that of their normal siblings. Our data suggested that excessive sleep fragmentation was a basic feature of narcolepsy and that it betrayed a constitutional predisposition for sleep to dissociate into its components and to become distributed around the nycthemeron. This process could be aggravated by emotional stress and by environmental demands for sustained vigilance, and this in turn, created the differences in symptoms and signs between individuals with identical genetic predispositions.

Aged↗
PubMed2001

The nature of collision-induced dissociation processes of doubly protonated peptides: comparative study for the future use of matrix-assisted laser desorption/ionization on a hybrid quadrupole time-of-flight mass spectrometer in proteomics.

Comparative MS/MS studies of singly and doubly charged electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) precursor peptide ions are described. The spectra from these experiments have been evaluated with particular emphasis on the data quality for subsequent data processing and protein/amino acid sequence identification. It is shown that, once peptide ions are formed by ESI or MALDI, their charge state, as well as the collision energy, is the main parameter determining the quality of collision-induced dissociation (CID) MS/MS fragmentation spectra of a given peptide. CID-MS/MS spectra of singly charged peptides obtained on a hybrid quadrupole orthogonal time-of-flight mass spectrometer resemble very closely spectra obtained by matrix-assisted laser desorption/ionization post-source decay time-of-flight mass spectrometry (MALDI-PSD-TOFMS). On the other hand, comparison of CID-MS/MS spectra of either singly or doubly charged ion species shows no dependence on whether ions have been formed by ESI or MALDI. This observation confirms that, at the time of precursor ion selection, further mass analysis is effectively decoupled from the desorption/ionization event. Since MALDI ions are predominantly formed as singly charged species and ESI ions as doubly charged, the associated difference in the spectral quality of MS/MS spectra as described here imposes direct consequences on data processing, database searching using ion fragmentation data, and de novo sequencing when ionization techniques are changed.

Animals↗
PubMed2001

Cryptic epitopes in N-terminally truncated prion protein are exposed in the full-length molecule: dependence of conformation on pH.

Prion diseases are diseases of protein conformation. Structure-dependent antibodies have been sought to probe conformations of the prion protein (PrP) resulting from environmental changes, such as differences in pH. Despite the absence of such antibodies for full-length PrP, a recombinant Fab (D13) and a Fab derived from mAb 3F4 showed pH-dependent reactivity toward epitopes within the N-terminus of N-terminally truncated PrP(90-231). Refolding and maintaining this protein at pH > or =5.2 before immobilization on an ELISA plate inhibited reactivity relative to protein exposed to pH < or =4.7. The reactivity was not affected by pH changes after immobilization, showing retention of conformation after binding to the plate surface, although guanidine hydrochloride at 1.5-2 M was able to expose the cryptic epitopes after immobilization at pH > or =5.2. The alpha-helical CD spectrum of PrP(90-231) refolded at pH 5.5 was reduced somewhat by these pH changes, with a minor shift toward beta-sheet at pH 4 and then toward coil at pH 2. No covalent changes were caused by the pH differences. This pH dependence suggests titration of an acidic region that might inhibit the N-terminal epitopes. A similar pH dependence for a monoclonal antibody reactive to the central region identified an acidic region incorporating Glu152 as a significant participant.

Amino Acid Sequence↗
PubMed1993

Tumor cell retention of antibody Fab fragments is enhanced by an attached HIV TAT protein-derived peptide.

Two peptide analogs of the 37-62 sequence region of the HIV TAT protein bind tightly to the surface of A431 breast carcinoma cells. After conjugation to either of two poorly internalized anti-tumor antibody Fab fragments, the analogs enhanced the in vitro cell surface retention and internalization of the Fab fragments to the level of the whole antibodies. This was at the expense of some binding specificity in the case of 1.6 peptides/NRLU-10 Fab, but not in the case of 1.1 peptides/Fab. Enhanced retention may occur by enhanced bivalent binding of the Fab fragments. The internalized fraction of free peptide, but not of the Fab conjugates, is enhanced by chloroquine. The conjugates which were less specific for tumor cell binding may be useful for enhanced retention/internalization of specifically acting agents, for use at specific sites of injection, or against pre-separated target cell populations, while the more specific conjugate may be of interest for further development.

Amino Acid Sequence↗