SEARCH · PubMed Health

Results for “Characterization”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 613 records · Page 34Linked to original sources

PubMed2001

Isolation of peptides from porcine intestinal tissue that induce extracellular acidification in CHO cells: identification of the active peptide as IGF-I and characterization of a fragment of calponin H1 processed at a dibasic site.

Chinese hamster ovary (CHO) cells are widely used as hosts for receptor expression and pharmacological studies. However, several endogenous receptor populations are present on these cells. Intestinal tissue extracts were found to induce strong extracellular acidification responses (ECAR) in CHO cells, yet several pure hormonal peptides, such as VIP, secretin, CCK, GIP, and galanin were ineffective. It is not known, which are the active compounds in the extracts that can stimulate the extracellular acidification in CHO cells. These active substances may be ligands for yet unknown receptors that are present natively in this cell type. We therefore decided to identify the active compound(s) by isolation from intestinal extract and structural characterization. Using chromatographic separations in combination with microphysiometry we have purified and characterized one such bioactive ligand. Structural analysis indicated that the isolated peptide was identical to insulin-like growth factor I (IGF-I). In the intestine, IGF-I is present in low amounts and has previously been detected only with radioimmunoassays. The results indicate that CHO cells express functional receptors for IGF-I. Among the peptides extracted from the intestine IGF-I is probably the strongest stimulator of ECAR in CHO cells. Moreover, IGF-I acts synergistically with other factors present in the crude tissue extract. Additionally, a fragment of calponin H1 (residues 1-43), previously not described at the protein level, was identified in the IGF-I containing fractions. The fragment was characterized by mass spectrometry and found to be N-terminally modified by acetylation suggesting that the whole protein bears the same posttranslational modification.

Acetylation↗

PubMed2001

Biophysical characterization of a soluble CD40 ligand (CD154) coiled-coil trimer: evidence of a reversible acid-denatured molten globule.

The CD40 ligand molecule is unique, consisting of a receptor-binding domain anchored by an isoleucine zipper moiety. Exact determination of the multimeric state and its tendency to form molten globules has not been elucidated. Corroborating evidence of a trimerized molecule in aqueous solution was obtained from size-exclusion chromatography, laser light scattering, and analytical ultracentrifugation. A reversible acid-denatured molten globule state was observed from circular dichroism and fluorescence spectroscopy data. The molten globule state was characterized by a loss of tertiary structure with associated retention of secondary structure near pH 3. Once returned to pH 7, the acid-denatured state refolded over the course of 7 days resulting in approximately 90% recovery of the native structure. The molten globule state was characterized by a broadening of structural features in the second-derivative spectra of Fourier transform infrared spectroscopy. A component band at 1650 cm(-1) was shown to be alpha-helix and originate from amide carbonyl vibrations of the isoleucine zipper. Differential scanning calorimetry measurements characterized the pH-sensitive molten globule state at pH 3.3 as one lacking a well-defined unfolding transition with an accompanying baseline shift at 58 degrees C (a consequence of increased heat capacity). The tendency to form molten globules during acid denaturation stress permits an opportunity to study the process of partial protein unfolding with implications concerning stability. Although reversible molten globules can be formed, it is important to recognize the unusual nature since the molten globule state is formed exclusively within the beta-sheet receptor-binding region.

Animals↗

PubMed1997

Use of a resonant mirror biosensor to characterize the interaction of carboxypeptidase A with an elicited monoclonal antibody.

The binding of apocarboxypeptidase A to an immobilized form of its elicited monoclonal antibody has been used to explore the potential of a biosensor instrument (IAsys) based on resonant mirror technology for the quantitative characterization of antibody-antigen interactions. Advantage has been taken of the stirred cuvette design of the IAsys instrument to develop a stepwise titration procedure for thermodynamic characterization of the interaction, an association equilibrium constant of 3.3 (+/-0.9) x 10(7) M-1 having been obtained under the conditions studied (0.1 M Tris/HCl-0.5 M NaCl, pH 7.5, 21 degrees C). In a test of the feasibility of subjecting the time course of biosensor response to conventional pseudo-first-order kinetic analysis, nonconformity of results with such description was encountered at high and low concentrations of apocarboxypeptidase A. Whereas the deviations from Langmuirian kinetic behavior at high antigen concentrations undoubtedly stem from the same sources as those already encountered in studies with the BIA-core biosensor instrument, the deviations at the other concentration extreme occur in a range in which the assumed constancy of free antigen concentration in the liquid phase is becoming a poor approximation. An alternative approach in such circumstances has been tested in which prior thermodynamic characterization is a prerequisite for rate constant evaluation by means of a second-order kinetic analysis. Finally, the effect of soluble anticarboxypeptidase A on the pseudo-first-order kinetics of the biosensor response has been used to illustrate a simple kinetic procedure for evaluating the affinity constant for the antibody-antigen interaction in solution, a value of 1.9 (+/-0.2) x 10(8) M-1 being obtained by such means.

Antibodies, Monoclonal↗

PubMed2002

Characterization of tetanus toxin, neat and in culture supernatant, by electrospray mass spectrometry.

A method was developed for the liquid chromatographic-mass spectrometric (LC-MS) identification of extremely neurotoxic toxins. The method combines sample treatment in a safety containment and analysis of detoxified material in a common laboratory facility. The method was applied to the characterization of neat tetanus toxin and subsequent identification of the toxin in cell lysate supernatants and culture supernatants from different Clostridium tetani bacteria strains. Characterization of the neat toxin was accomplished by (1) accurate mass measurement of enzyme digest fragments of the toxin and (2) tandem mass spectrometric (MS/MS) amino acid sequencing of selected peptides. Accurate mass measurement proved no longer feasible for the analysis of supernatants, due to the overwhelming presence of peptides from proteins other than toxin. Even when high-molecular-weight proteins were filtered from the lysates and treated, the retained protein fraction yielded too many peptides. However, MS/MS could successfully be applied when the findings from the characterization of neat toxin were employed. Thus, LC-MS/MS of selected precursor ions from trypsin digest fragments yielded specific sequence data for identification of the toxin. This procedure provided reliable identification of the toxin at levels above 1 microg/ml and within a day. Investigations with the method developed will be extended to the botulinum neurotoxins.

Amino Acid Sequence↗

PubMed1993

Purification and characterization of a scorpion defensin, a 4kDa antibacterial peptide presenting structural similarities with insect defensins and scorpion toxins.

Insect defensins are a group of inducible small-sized antibacterial peptides with three intramolecular disulfide bridges. NMR studies have recently shown that they share striking structural similarities with scorpion toxins. We have investigated in a scorpion species, Leiurus quinquestriatus, the potential presence of antibacterial molecules and report the isolation and structural characterization of a novel insect defensin homologue, which we refer to as scorpion defensin. This peptide shows a remarkably high degree of sequence homology with a defensin recently characterized in a species belonging to the ancient insect order of the Odonata with which it defines a novel ancient subclass of defensins. The scorpion defensin has in common with the scorpion toxins a consensus sequence Cys-[...]-Cys-Xaa-Xaa-Xaa-Cys-[...]-Gly-Xaa-Cys-[...]-Cys-Xaa-Cys present in all scorpion toxins characterized so far.

Amino Acid Sequence↗

PubMed1993

Characterization of murine E-selectin expression in vitro using novel anti-mouse E-selectin monoclonal antibodies.

A recombinant phage containing the structural exons for mouse E-selectin has been isolated and characterized. Utilizing PCR techniques the lectin and egf domains were fused to form an artificial cDNA for expression in eukaryotic cells. Transient expression in COS cells demonstrated the lectin and egf domains were sufficient to mediate the binding of mouse and human neutrophils as well as HL60 cells. Recombinant soluble mouse E-selectin was purified and used to immunize rats to generate mAbs specific to mouse E-selectin. A panel of mAbs directed against mouse E-selectin was characterized including five that inhibit the adhesion of HL60 cells or mouse neutrophils to COS cells expressing the mouse lectin/egf domains. These mAbs have been used to characterize the expression and function of E-selectin on cytokine stimulated eEnd.2 murine endothelial cells.

Animals↗

PubMed1994

Characterization of activin A-, activin AB- and activin B-responding cells by their responses to hypothalamic releasing hormones.

Activin A-, AB- and B-responding cells were characterized by their responsiveness in cytosolic free calcium ([Ca2+])i) to four hypothalamic releasing hormones, CRH, GHRH, TRH and GnRH. First, rat pituitary cells responding to activin A, AB and B in [Ca2+]i were determined in a mixed population of pituitary cells. The populations of the activin A-, AB-, and B-responding cells were 13.9%, 9.3% and 13.2%, respectively. Overlapping of response among each population of activin- responding cells was present in some of the cells. The cells responding to activin A, AB, and B were then characterized by their responses to CRH, GHRH, TRH and GnRH. Most of the cells responding to activin A, AB, and B also responded to GHRH or TRH. These results reveal that there are distinct differences among each population of activin A-, AB- and B-responding cells and that there is still functional overlapping of responsiveness among these populations. The characterization of activin-responding cells suggests involvement of somatotropes and lactotropes in activin-induced biological events in the pituitary.

Activins↗

PubMed1995

Detection and characterization of apoptosis in osteoclasts in vitro.

Morphological changes of nuclei and DNA fragmentation undergoing cell death were studied using highly purified rabbit osteoclasts in vitro. Approximately fifty percent of dead osteoclasts exhibited morphological changes of nuclei characterized by apoptosis and approximately 70% of osteoclasts with changes of nuclei exhibited DNA fragmentation in our culture system. These changes of nuclei were confirmed as condensation of chromatin through transmission electronmicroscope. Moreover, biochemical characterizations of these changes were compatible with the characterizations of apoptosis in other cell systems. These results demonstrate that part of osteoclasts die by a mode of apoptosis.

Animals↗

PubMed1997

Characterization of gamma S-crystallin isoforms from lip shark (Chiloscyllium colax): evolutionary comparison between gamma S and beta/gamma crystallins.

gamma S-Crystallin from shark eye lenses, formerly termed beta s crystallin in mammalian lenses, is structurally characterized in this study by cDNA cloning and sequencing. To facilitate sequence characterization of gamma S-crystallin possessing intermediate structural properties between beta- and gamma-crystallins, cDNA mixture was constructed from the poly(A)+ mRNA isolated from shark eye lenses, and amplification by polymerase chain reaction (PCR) was carried out to obtain nucleotide segments encoding multiple shark gamma S-crystallins. Sequencing several positive clones revealed that a multiplicity of isoforms exists in the gamma S-crystallin class of this cartilaginous fish, similar to authentic gamma-crystallin family characterized from the same shark species. Comparison of protein sequences encoded by two representative shark gamma S1 and gamma S2 cDNAs with those published sequences of beta-, gamma-, and gamma S crystallins from bovine, human, bullfrog and carp lenses indicated that there is about 35-64% sequence homology between shark gamma S crystallins and structurally related crystallins from different evolutionary classes, with a higher sequence similarity between shark gamma S and mammalian gamma-crystallins than that of shark gamma S and carp gamma S or bovine gamma S crystallins. A phylogenetic tree constructed on the basis of the sequence divergence among various beta-, gamma-, and gamma S crystallins corroborates the closer relatedness of shark gamma S to authentic gamma-crystallin than to mammalian and teleostean gamma S crystallins. It further strengthens the supposition that ancestral precursors of gamma S-crystallins were present in the shark lens long before the appearance of present-day teleostean and mammalian gamma S-crystallins.

Amino Acid Sequence↗

PubMed1998

Characterization of gammaS-crystallin isoforms from a catfish: evolutionary comparison of various gamma-, gammaS-, and beta-crystallins.

gammaS-Crystallin from catfish eye lenses, formerly designated betas-crystallin in mammalian lenses, is structurally characterized in this study by cDNA cloning and sequencing. To facilitate sequence characterization of gammaS-crystallin with structural properties lying between beta- and gamma-crystallins, a cDNA mixture was constructed from the poly(A)+ mRNA isolated from catfish eye lenses, and amplification by polymerase chain reaction (PCR) was carried out to obtain nucleotide segments encoding multiple gammaS-crystallin isoforms. Sequencing several positive clones revealed that at least two distinct isoforms exist in the gammaS-crystallin class of this teleostean fish, similar to the authentic gamma-crystallin family characterized previously in species of the piscine class. Comparison of protein sequences encoded by two representative catfish gammaS1 and gammaS2 cDNAs with the published sequences of beta-, gamma-, and gammaS-crystallins from shark, carp, bullfrog, bovine, and human lenses indicates that there is about 20-50% sequence homology between catfish gammaS-crystallins and various members of the related beta/gamma-crystallin superfamily from different evolutionary classes, with a higher sequence similarity being found between catfish gammaS- and mammalian gamma-crystallins than between catfish gammaS- and bovine or carp gammaS-crystallins. Phylogenetic trees constructed on the basis of the nucleotide and protein sequence divergence among various beta-, gamma-, and gammaS-crystallins corroborate the closer relatedness of catfish gammaS- to authentic gamma-crystallin than to bovine and carp gammaS-crystallins. The results suggest that evolution of catfish gammaS-crystallins follows a different path from that of bovine and carp gammaS-crystallins and may represent a more ancient offshoot from the ancestral gamma/gammaS coding gene than carp and bovine gammaS-crystallins.

Amino Acid Sequence↗

PubMed2000

Isolation and characterization of the human UGT2B7 gene.

Glucuronidation is a major pathway involved in the metabolism of drugs and numerous endogenous compounds, such as bile acids and steroid hormones. The enzymes responsible for this conjugation reaction are UDP-glucuronosyltransferases (UGT). Among the UGT2B subfamily, UGT2B7, a UGT enzyme present in the liver and several steroid target tissues, is an important member since it conjugates a large variety of compounds including estrogens, androgens, morphine, AZT, and retinoic acid. Although this enzyme is well characterized, the gene encoding the UGT2B7 protein and its promoter region remain unknown. In this article, we report the genomic organization and the promoter region of the human UGT2B7 gene. To isolate this gene, a P-1 artificial chromosome (PAC) library was screened with a full length UGT2B7 probe and a clone of approximately 100 kb in length was isolated. In addition to the UGT2B7 gene, this PAC contains two other UGT2B genes previously characterized, namely UGT2B26P and UGT2B27P. The UGT2B7 gene is composed of six exons spanning approximately 16 kb, with introns ranging from 0.7 to 4.2 kb. The 5'-flanking region of the human UGT2B7 gene contains several potential cis-acting elements such as Oct-1, Pbx-1, and C/EBP. Only one TATA-box at nucleotide -106 was found within the first 500 nucleotides relative to the adenine base of the initiator ATG codon. Characterization of the UGT2B7 gene provides insight into the organization and regulation of this important metabolic gene.

Base Sequence↗

PubMed2001

Recombinant expression, purification, and kinetic characterization of chondroitinase AC and chondroitinase B from Flavobacterium heparinum.

Glycosaminoglycans (GAGs) are a family of complex polysaccharides involved in a diversity of biological processes, ranging from cell signaling to blood coagulation. Chondroitin sulfate (CS) and dermatan sulfate (DS) comprise a biologically important subset of GAGs. Two of the important lyases that degrade CS/DS, chondroitinase AC (EC 4.2.2.5) and chondroitinase B (no EC number), have been isolated and cloned from Flavobacterium heparinum. In this study, we outline an improved methodology for the recombinant expression and purification of these chondroitinases, thus enabling the functional characterization of the recombinant form of the enzymes for the first time. Utilizing an N-terminal 6x histidine tag, the recombinant chondroitinases were produced by two unique expression systems, each of which can be purified to homogeneity in a single chromatographic step. The products of exhaustive digestion of chondroitin-4SO(4) and chondroitin-6SO(4) with chondroitinase AC and dermatan sulfate with chondroitinase B were analyzed by strong-anion exchange chromatography and a novel reverse-polarity capillary electrophoretic technique. In addition, the Michaelis-Menten parameters were determined for these enzymes. With chondroitin-4SO(4) as the substrate, the recombinantly expressed chondroitinase AC has a K(m) of 0.8 microM and a k(cat) of 234 s(-1). This is the first report of kinetic parameters for chondroitinase AC with this substrate. With chondroitin-6SO(4) as the substrate, the enzyme has a K(m) of 0.6 microM and a k(cat) of 480 s(-1). Recombinantly expressed chondroitinase B has a K(m) of 4.6 microM and a k(cat) of 190 s(-1) for dermatan sulfate as its substrate. Efficient recombinant expression of the chondroitinases will facilitate the structure-function characterization of these enzymes and allow for the development of the chondroitinases as enzymatic tools for the fine characterization and sequencing of CS/DS.

Bacterial Proteins↗

PubMed1995

Characterization of murine brain-reactive monoclonal IgG autoantibodies.

A diversity of brain-reactive autoantibodies (BRAA) is found in the sera and cerebrospinal fluid in elevated amounts in systemic lupus erythematosus (SLE) and correlates with some CNS manifestations of both human and murine SLE. In order to test the hypothesis that BRAA mediate the CNA manifestations, we are developing a library of brain-reactive monoclonal autoantibodies (BRMA) from autoimmune mice for characterization. We have recently reported on the production of BRMA of the IgM class from unimmunized autoimmune mice. In the current study we extend our investigations and report on the production and characterization of two BRMA of the IgG class. These antibodies react against integral brain membrane antigens of approximately 58, 32, and 30 kDa by immunoblot. Reactivity to such antigens is also found in a majority of the autoimmune murine serum samples tested, but not in nonautoimmune mice. These IgG BRMA show reactivity to cell bodies of the cerebral cortex, hippocampus, and hypothalamus of murine brain but not to fiber tracts. They also react with an integral thymus membrane antigen, but not to antigens of other tissues tested. Because of their properties, BRMA such as those characterized here are likely to be of pathogenic significance in CNS involvement in SLE.

Animals↗

PubMed2001

Application of Bayesian inference to characterize risks associated with low doses of low-LET radiation.

Improved risk characterization for stochastic biological effects of low doses of low-LET radiation is important for protecting nuclear workers and the public from harm from radiation exposure. Here we present a Bayesian approach to characterize risks of stochastic effects from low doses of low-LET radiation. The stochastic effect considered is neoplastic transformation of cells because it relates closely to cancer induction. We have used a published model of neoplastic transformation called NEOTRANS1. It is based on two different classes of cellular sensitivity for asynchronous, exponentially growing populations (in vitro). One sensitivity class is the hypersensitive cell; the other is the resistant cell. NEOTRANS1 includes the effects of genomic damage accumulation, DNA repair during cell cycle arrest, and DNA misrepair (non-lethal repair errors). The model-associated differential equations are solved for conditions of in vitro irradiation at a fixed rate. Previously published solutions apply only to high dose rates and were incorrectly assumed to apply to only high-LET radiation. Solutions provided here apply to any fixed dose rate and to both high- and low-LET radiations. Markov chain Monte Carlo methods are used to carry out the Bayesian inference of the low-dose risk for neoplastic transformation of aneuploid C3H 10T1/2 cells for X-ray doses from 0 to 1000 mGy. We have assumed that for this low-dose range only the hypersensitive fraction of the cells are affected. Our results indicate that the initial slope of the risk vs dose relationship for neoplastic transformation is as follows: (1) directly proportional to the fraction, f1, of hypersensitive cells; (2) directly proportional to the radiosensitivity of the genomic target; and (3) inversely proportional to the rate at which hypersensitive cells with radiation-induced damage are committed to undergo correct repair of genomic damage. Further, our results indicate that very fast molecular events are associated with the commitment of cells to the correct repair pathway. Results also indicate a relatively large probability for misrepair that leads to genomic instability. Our results are consistent with the view that for very low doses, dose rate is not an important variable for characterizing low-LET radiation risks so long as age-related changes in sensitivity do not occur during irradiation.

Animals↗

PubMed1995

Characterization of a novel human antibody xenoreactive with fibronectin.

Recent reports have used bovine fibronectin (Fn) as source of antigen to study human anti-Fn autoantibodies. We have characterized a novel human antibody (Ab) reactive with bovine and marsupial Fn, but not with human Fn. Indirect immunofluorescence, wet cleaving and protein adherence assays, immunoblotting, blot-affinity purification, a cell adhesion inhibition assay, and competitive experiments with synthetic peptides were used to characterize the anti-Fn Ab in serum from a patient with an undifferentiated connective tissue disease. A characteristic Fn-like network was observed by indirect immunofluorescence on bovine MDBK and marsupial PtK2 cells, but not on various human cell lines. Double immunofluorescence revealed colocalization of the Ab with a mouse monoclonal anti-Fn Ab. A reactive polypeptide of 240 kDa corresponding to the M(r) of Fn was identified by immunoblotting using MDBK and PtK2 total cell lysates. The Ab reacted with the 240-kDa band of purified bovine Fn with an endpoint titer of 1:64,000, while no reactivity was observed with human cellular or plasma Fn. Blot-affinity purification of the Ab from the 240-kDa PtK2 region confirmed that the Fn-like fluorescent pattern observed was due to reactivity with the 240-kDa band and not with other regions of the blot. The Ab affinity-purified from the 240-kDa region also reacted with purified bovine Fn by immunoblotting. Functional analysis disclosed specific inhibition of PtK2 and MDBK cell adhesion by the affinity-purified anti-Fn Ab. Competitive experiments with synthetic peptides demonstrated that the epitope is located in the decapeptide RGDSPASSKP containing the cell-binding domain of Fn. Longitudinal analysis of the Ab revealed its persistence over 6 years. Bovine and marsupial Fn can be the focus of a highly specific and persistent human immune response. Reactivity of a human Ab with bovine Fn does not imply cross-reactivity with human Fn. In light of recent reports using bovine Fn to characterize human anti-Fn "autoantibodies," future studies on human anti-Fn should specifically employ purified human Fn as antigen.

Adult↗

PubMed2001

Toxoplasma gondii: molecular cloning and characterization of a novel 18-kDa secretory antigen, TgMIC10.

Hoff, E. F., Cook, S. H., Sherman, G. D., Harper, J. M., Ferguson, D. J. P., Dubremetz, J. F., and Carruthers, V. B. 2001. Toxoplasma gondii: Molecular cloning and characterization of a novel 18-kDa secretory antigen, TgMIC10. Experimental Parasitology, 97, 77-88. During host cell invasion, Toxoplasma gondii secretes proteins from specialized organelles (micronemes and rhoptries) located at the apical end of the parasite. The contents of the micronemes appear to be crucial to T. gondii invasion, as inhibition of microneme secretion prevents parasite entry into host cells. Here we describe a new T. gondii microneme protein, TgMIC10. Molecular characterization of a full-length TgMIC10 cDNA revealed that TgMIC10 lacks homology to any previously characterized proteins, although a homologue, NcMIC10, was identified in a closely related parasite, Neospora caninum. TgMIC10 has an unusually long secretory leader sequence of 58 amino acids; the mature TgMIC10 is 18 kDa, possesses nine diglutamic acid repeats and an imperfect repeat sequence (RK(R/Y)HEEL), and is entirely devoid of cysteines. Antibodies raised against recombinant TgMIC10 recognized the native TgMIC10 and localized the protein to the micronemes in indirect immunofluorescence and immunoEM experiments. Comparison of immunofluorescence images indicates that TgMIC10 expression is higher in T. gondii tachyzoites, which are responsible for active infection, than in bradyzoites, which are responsible for latent infection.

Amino Acid Sequence↗

PubMed1997

Characterization of human SHC p66 cDNA and its processed pseudogene mapping to Xq12-q13.1.

SHC is an adapter protein in the Ras-MAPkinase pathway that is involved in the regulation of cell growth and differentiation. The p46 and p52 isoforms are thought to be produced by the use of two alternative translation initiation sites in a 3.4-kb transcript from the SHCA gene, which maps to chromosome 1q21. The p66 isoform could be encoded by a different 3.8- or 2.8-kb transcript of the same gene or alternatively by a SHC-related gene. To characterize other putative genes coding for SHC-like proteins, primers from the 3' UTR of the SHCA gene were used to screen a yeast artificial chromosome (YAC) library by polymerase chain reaction (PCR). Two YAC clones, 20D11B and 36D1D, were isolated and used as probes for fluorescence in situ hybridization analysis. Both these probes hybridized to chromosome Xq12-q13.1. This novel SHC-related sequence was characterized by direct sequencing of vectorette library PCR products produced from clone 20D11B. A transcript of 3.2 kb that was 85% identical to the mouse Shc cDNA encoding the p66 isoform was identified. Sequence analysis demonstrated the presence of multiple stop codons identifying this isoform of SHC as a processed pseudogene. Using primers designed on the basis of the nucleotide sequence of the pseudogene, we have now amplified and sequenced a human cDNA that encodes the SHC p66 protein. Thus, we have characterized the human SHC p66 isoform cDNA and identified a processed SHC pseudogene that maps to chromosome Xq12-q13.1.

Adaptor Proteins, Signal Transducing↗

PubMed1998

Cloning and characterization of the multiple murine homologues of NAIP (neuronal apoptosis inhibitory protein).

The spinal muscular atrophies (SMAs), characterized by degeneration of spinal cord motor neurons, are among the most common autosomal recessive disorders. We have previously reported the characterization of an SMA-associated gene designated NAIP (neuronal apoptosis inhibitory protein). This gene, which encodes a protein homologous to the baculoviral inhibitor of apoptosis proteins, is deleted in a significant proportion of individuals with type I SMA, is expressed in motor neurons, and inhibits apoptosis both in vitro and in vivo. Here we present the cloning and characterization of multiple copies of the mouse homologue of NAIP, Naip1-Naip6. Our analysis of the genomic organization of Naip indicated the existence of a minimum of six distinct Naip loci in the 129/SvJ mouse strain. However, Southern blot analysis revealed that only three of these loci contained the 5'UTR element essential for translation in the CNS. The coding region of one of these three potentially functional loci (Naip1) demonstrates 77% homology to NAIP at the nucleotide level and 68% identity at the amino acid level.

Animals↗