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PubMed1993

Taxonomy of lactic acid bacteria from spoiled, vacuum packaged vienna sausages by total soluble protein profiles.

To resolve the taxonomic status of sixty-one lactic acid bacteria isolated from spoiled, vacuum packaged vienna sausages, a computer assisted analysis of total soluble protein profiles was undertaken. Sixty-two% of the strains in the study were recovered in fourteen clusters at the 72% similarity level, indicating high degree of heterogeneity among the strains examined. Two clusters representing Leuconostoc species were the largest and most homogeneous, containing twenty strains between them. One cluster contained both typical Lactobacillus saké and Lactobacillus curvatus strains, indicating the inability of protein profiles to consistently differentiate between all strains of these two species. The presence of an independent Lactobacillus curvatus grouping and the clustering of some atypical strains with Lactobacillus saké isolates, however, indicated that the atypical strains may be closely related to Lactobacillus saké.

Bacterial Proteins↗

PubMed1996

Serum insulin-like growth factor binding protein profiles in postmenopausal women: their correlation with bone mineral density.

OBJECTIVE: The objective of this study was to evaluate the relationship between serum insulin-like growth factor binding protein profiles and bone mineral density measurements and biochemical markers of bone turnover in postmenopausal women. STUDY DESIGN: Sera from 37 normal healthy postmenopausal women and 14 postmenopausal women with osteoporosis were analyzed for insulin-like growth factor binding proteins and osteocalcin by Western ligand blotting and radioimmunoassay, respectively. Bone mineral density measurements were performed by dual-energy x-ray absorptiometry. Urinary calcium and creatinine excretion were also determined. The data were analyzed statistically by analysis of variance and the least-squares method. RESULTS: The serum insulin-like growth factor binding protein-2 ratio (insulin-like growth factor binding protein-2 intensity/total insulin-like growth factor binding protein intensity) in postmenopausal women with osteoporosis was significantly higher (p < 0.02) than that in normal healthy postmenopausal women, but the serum insulin-like growth factor binding protein-3 ratio in women with osteoporosis was significantly lower (p < 0.01). The serum insulin-like growth factor binding protein-2 ratio correlated negatively with the bone mineral density of the lumbar spine (p < 0.0001), femoral neck (p < 0.05), trochanter (p < 0.01), and Ward's triangle (p < 0.05), but there was a significant positive correlation between the serum insulin-like growth factor binding protein-3 ratio and the bone mineral density of the lumbar spine (p < 0.001) and trochanter (p < 0.05). No correlations between serum insulin-like growth factor binding protein levels or serum ratios and biochemical markers of bone turnover were noted. CONCLUSION: The measurement of serum insulin-like growth factor binding protein profiles might be useful in identifying postmenopausal women at risk for osteoporosis.

Bone Density↗

PubMed1976

Lipid composition and protein profiles of outer and inner membranes from pig heart mitochondria. Comparison with microsomes.

1. Mitochondria, inner and outer mitochondrial membranes and microsomes were isolated and purified from pig heart. Their lipid composition and protein components were studied. 2. The fatty acid distribution in the main phospholipids seemed specific rather of a given phospholipid and not of one type of membrane. 3. Inner mitochondrial membranes were characterized by a high content in cardiolipin and a very low level of triglycerides together with a high degree of unsaturation and C18 acids. Gel electrophoresis revealed 13 different polypeptide subunits of which 5 were major ranging in molecular weights from 10000 to 215000. 4. In outer mitochondrial membranes, total lipid, phosphatidylcholine, phosphatidylinositol, plasmologen and triglyceride contents were much higher than in inner membranes. Fatty acids of phospholipids were mostly saturated and the polypeptide pattern showed 12 components, of which 4 were major of mol. wt 75000, 60000, 20000 and below 10000. 5. Compared to outer membrane, microsomes exhibited a much higher cholesterol content and markedly different protein profiles. They contained significant amounts of cardiolipin and phosphatidylserine, this latter phospholipid being exclusively located in microsomes. However odd similarities were observed in some lipid components of microsomes and inner mitochondrial membranes, but fatty acids were more saturated in microsomes and electrophoretic profiles of protein components appeared very different and revealed components of high mol. wt.

Animals↗

PubMed2000

[Protein profile of Senegalese children presenting with nephrotic syndrome].

In Senegal, nephrotic syndrome usually occurs upon malnutrition that confers it its specificity and has brought us to establish the proteinic profile of Senegalese children suffering from it, in the aim to improve its treatment. Twenty children on the average of six years, hospitalized for nephrotic syndrome, were chosen. They were compared to 20 healthy children of the same age group, and 20 other children hospitalized for a non-renal inflammatory syndrome. For each subject were dosed total blood proteins and albumin. The protein electrophoresis has permitted to evaluate the alpha-1, alpha-2, beta and gamma globulins. We have observed predominance of boys with a 2.33 sex ratio. Our patients presenting a nephrotic syndrome were about six years old; no case has been noted before two years. Total proteins and albumin have decreased respectively to 52.10 g/l and 16 g/l. This diminishing is essentially linked to the massive urinary escape of plasmatic proteins and would explain partly the increase of lipids. The alpha-1 globulins were equally lowered but the alpha-2 globulins were increased by 40% of total proteins. Beta globulins have decreased, following the evolution of gamma globulins. In Senegal, The early age of nephrotic syndrome occurrence and the biological perturbations of the proteinic profile, recommend global and quick treatment considering food recovery and nutrition. The correction of these parameters being able to constitute in one hand an element of accessible and reliable supervision, and one the other hand the basis of complete treatment.

Adolescent↗

PubMedDate not supplied

Urinary protein profiles following urography with iothalamate. A preliminary report.

Urinary protein profiles (IgG, albumin, beta 2-microglobulin) following urography with iothalamate were investigated in 5 patients with functionally and morphologically normal kidneys. In 4 of 5 patients an increased clearance of at least one of the proteins was found indicating disturbance in both glomerular and tubular function and the change was transient (up to 6 days). Albustix was always negative.

Albuminuria↗

PubMed1997

Altered nuclear matrix protein profiles in oncogene-transformed mouse fibroblasts exhibiting high metastatic potential.

The nuclear matrix provides the structural support of the nucleus and is involved in various cellular functions of the nucleus. Nuclear matrix proteins, which are both tissue and cell type specific, are altered with transformation and state of differentiation. Here, nuclear matrix protein profiles of oncogene-transformed mouse fibroblasts with various degrees of metastatic activity were analyzed using two-dimensional gel electrophoresis. This study shows that as the metastatic potential increases, similar nuclear matrix protein profiles are associated with each increase regardless of transformation agent.

3T3 Cells↗

PubMed1998

[Non-Hodgkin lymphoma in adults. Protein profile of CSF and serum in 25 patients].

Twenty-five non-Hodgkin's adult patients of a cohort studied for detection of neurologic involvement were evaluated on the cerebrospinal fluid (CSF) protein profile. CSF and serum were collected in the same occasion. Blood-brain barrier and local synthesis of IgG were studied. There was an incidence of neurologic signs and symptoms in 48% of all patients. Samples analysis showed: increase of total protein in CSF in 52%; local synthesis of IgG in one HIV seropositive patient; IgG concentration increase in the CSF in the absence of malignant cells in the CSF in two patients that clinically improved after chemotherapy; oligoclonal bands only in the CSF in one HTLV-I seropositive patient. These data show that the study of CSF protein profile can contribute in the characterization of CNS involvement in non-Hodgkin lymphoma.

Adult↗

PubMed2004

A combination of protein profiling and isotopomer analysis using matrix-assisted laser desorption/ionization-time of flight mass spectrometry reveals an active metabolism of the extracellular matrix of 3T3-L1 adipocytes.

Differential gel electrophoresis followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is a commonly used protein profiling method. However, observed changes can be explained in multiple ways, one of which is by the protein turnover rate. In order to easily and rapidly obtain information on both the identity and turnover of individual proteins, we applied a combination of protein labeling with L-(ring-2,3,4,5,6 2H5) phenylalanine and MALDI-TOF MS. While the spectrum reveals the identity of a protein, mass isotopomer analysis provides information about the rate of protein labeling as a measure of synthesis or turnover. Using this approach on mature 3T3-L1 adipocytes, we were able to discriminate between rapidly and slowly metabolised proteins. In our isolate, proteins of the cytoskeleton appeared to be slowly metabolised, whereas components of the extracellular matrix, in particular collagen type I alpha 1 (COL1A1) and collagen type I alpha 2 (COL1A2) showed rapid accumulation of newly synthesized proteins. Both proteins appeared to be metabolised in the same ratio as they are present in collagen fibers, i.e. 2:1 (COL1A1: COL1A2). In addition, functionally related proteins were also readily labeled. Taken together, we have shown that a combination of stable isotope labeling and protein profiling by gel electrophoresis and MALDI-TOF analysis can simultaneously provide information on the identity and relative metabolic rate of proteins in eukaryotic cells in a simple, nonhazardous and rapid-throughput way.

3T3-L1 Cells↗

PubMed1986

Comparative studies on the protein profiles and hydrophobicity of strains of Streptococcus mutans serotype c.

Twelve strains of Streptococcus mutans serotype c were grown in batch culture with glucose at constant pH (6.0) and a number of properties compared. On the basis of their cellular and extracellular protein profiles, the strains were divided into three groups, I, II and III, containing five, four and three strains, respectively. The extracellular protein profiles for a particular strain differed if the organisms were grown either at pH 6.0 with fructose instead of glucose or with glucose but without pH control. The total amount of extracellular protein produced by group III strains grown in glucose-containing medium at pH 6.0 was several times that produced by strains of groups I and II, which were also more hydrophobic. One of the potentially important proteins is P1, also called antigen B or I/II, and it was shown to be entirely in the culture fluid of group III strains but mostly cell-associated from strains of groups I and II. Approximately half of the cell-associated fraction of P1 could be removed with hot sodium dodecyl sulphate.

Bacterial Proteins↗

PubMed2006

Comparison between Holstein cow's milk and Japanese-Saanen goat's milk in fatty acid composition, lipid digestibility and protein profile.

The fatty acid composition, the lipid digestibility and protein profile of Japanese-Saanen goat's milk were characterized. Caprine milk contained substantial quantities of C(4:0) to C(10:0) fatty acids as compared with Holstein cow's milk. The lipids of the former showed significantly higher digestibility in vitro by porcine lipase than those of the latter (P<0.05). As determined by SDS-PAGE, the respective contents of alpha(s1)-casein, one of the major allergens, were 3.9% and 33.7% in caprine and bovine milk.

Animals↗

PubMed1985

Human pancreatic secretory protein profiles in pancreas cancer and chronic pancreatitis.

Pancreatic secretory protein profiles differed significantly between patients with chronic pancreatitis (CP) and pancreatic carcinoma (CA). Specific regions of the patterns were altered when CP versus CA and when CP versus normals were compared. Bands isoelectric in the region of pH 9-11 were elevated in CP. The possible identification of this band as lactoferrin is discussed.

Chronic Disease↗

PubMed2004

[Establishment of protein profile of human small cell lung cancer cell line NCI-H446].

BACKGROUND & OBJECTIVE: Small cell lung cancer (SCLC) is particularly aggressive, and characterized by rapid growth and early metastasis. At present, there is no data concerning SCLC two-dimensional polyacrylamide gel electrophoresis (2-DE) reference map,and its protein profiles in public databases. This study was to establish a well-resolved, reproducible 2-DE map of proteome in SCLC cell line NCI-H446, and analyze its protein profiles. METHODS: Two-DE was applied to separate the total proteins of NCI-H446 cells, which were then silver-stained in the gel. Well-separated protein spots were selected from the gel by ImageMaster 2D analysis system. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), peptide map fingerprinting (PMF),and database searching were used to identify the protein spots. RESULTS: Clear,well-resolved, reproducible 2-DE patterns of proteome in NCI-H446 cells were obtained. The average protein spots of 3 gels were 1506+/-74; and 1412+/-56 spots were matched with an average matching rate of 93.4%. The average deviation of spot position was (0.96+/-0.27) mm in IEF direction, and (1.24+/-0.41) mm in SDS-PAGE direction indicating relatively good reproducibility of the protein spots. Fifty-eight proteins were identified, certain proteins were products of oncogenes, and others were involved in cell cycle regulation, and signal transduction. CONCLUSIONS: A reference map of NCI-H446 cells was established,certain proteins were identified by MALDI-TOF-MS and PMF. These data will be useful for establishing human SCLC proteome database.

Amino Acid Sequence↗

PubMedDate not supplied

[Evaluation of different liquid media for the culture of Gardnerella vaginalis and characterization of the protein profile with PAGE-SDS electrophoresis].

3 liquid culture media: triptose soya broth supplemented with equine serum (TSB-es)/without supplement (TSB) and Casman broth (CB) were evaluated under different incubation conditions at 10% of CO2 and under aerobic conditions. There were no significant differences in the values of optic density for the TSB-es and CB media. However, on establishing the protein profile by PAGE-SDS for the evaluated conditions, the presence of protein of 14 kDa and the values of optic density allowed the selection of the CB medium for the culture of Gardnerella vaginalis. The sensitivity to different pH in this medium was studied. pH 7.2 proved to be optimum and it was established its growth cynetics. The analysis by PAGE-SDS of the complete cells corresponding to different clinical isolates of G. vaginalis with biochemical variations made possible to characterize the protein profile in this medium and showed similarity between the isolates.

Bacterial Proteins↗

PubMed2000

pH-dependent protein profiles of Helicobacter pylori analyzed by two-dimensional gels.

BACKGROUND: Helicobacter pylori survives transient exposure to extreme acid prior to adherence and growth on the gastric epithelium at neutral pH. MATERIALS AND METHODS: The effect of pH stress on protein profiles of H. pylori was observed using two-dimensional gel electrophoresis (2-D gels). H. pylori 26695 was grown microaerobically in tryptone-yeast extract broth, 3% fetal bovine serum. Growth in acid alkalinized the medium, whereas growth in base caused acidification. For 2-D gel analysis of protein profiles, cultures were grown in media buffered at pH 5.7 and at pH 7.5. RESULTS: Under all pH conditions, the most abundant proteins observed were the urease structural subunit UreB and the chaperonin GroEL. Growth in acid significantly increased the abundance of UreB. Thus, urease expression is not completely constitutive, as reported previously, but shows regulation by pH. Another protein observed only at low pH was identified as mammalian apolipoprotein A-I, possibly taken up by H. pylori from bovine serum in the growth medium. This finding, if confirmed, suggests that uptake of high-density lipoprotein from the human host may facilitate acquisition of cholesterol, required for formation of the unique cholesteryl glucosides in the membrane of H. pylori. In growth above pH 7, three stress proteins were induced: GroES (HspA), GroEL (HspB), and the antioxidant AhpC homolog TsaA. In addition, N-terminal sequence analysis identified five additional proteins that had not previously been reported on 2-D gels of H. pylori (FMN, SodB, TrxB, TsaA, and Tsr). CONCLUSIONS: In summary, our 2-D gel study reveals expression of several proteins dependent on growth pH.

Bacterial Proteins↗

PubMed2003

Discovery of distinct protein profiles specific for lung tumors and pre-malignant lung lesions by SELDI mass spectrometry.

OBJECTIVES: Early lung cancer detection and treatment remain a challenge. The efficacy of surface-enhanced laser desorption/ionization (SELDI) technology in lung cancer detection, has not been defined. This study identifies specific protein peak patterns in malignant lung tumors, and in pre-malignant airways epithelium showing neoplastic transformation. METHODS: Lung tumor specimens taken from patients participating in a lung cancer screening study (H. Lee Moffitt Cancer Center, Tampa, FL) were laser capture microdissected to obtain pure cell populations from frozen sections of normal lung, atypical adenomatous hyperplasia (AAH) and malignant tumors. SELDI mass spectrometry was used to generate protein profiles in each epithelial cell type. RESULTS: SELDI mass spectroscopy is highly reproducible in detecting lung tumor-specific protein profiles. Three peaks at 17-23 kDa mass range from tumor cells showed markedly increased compared with normal cells. The peak at 17250 Da was not detected in any of the normal cells. This peak appeared to be present at low levels in the atypical cell samples. CONCLUSIONS: This study demonstrates the feasibility of detecting "malignant" protein signatures from lung tumor and pre-malignant pulmonary epithelium using SELDI mass spectrometry. Although additional study is necessary to validate these patterns as unique diagnostic tools, these "malignant" protein signatures lend themselves to identification of populations at high-risk for lung cancer and for monitoring response to lung cancer chemopreventive agents.

Carcinoma, Non-Small-Cell Lung↗

PubMed1999

Rapid screening of protein profiles of human breast cancer cell lines using non-porous reversed-phase high performance liquid chromatography separation with matrix-assisted laser desorption/ionization time-of-flight mass spectral analysis.

Non-porous reversed-phase (NP-RP) HPLC has been used to rapidly generate protein profiles of whole cell lysates of human breast cancer cell lines. The non-porous packing material used was silica coated with C18, which provided rapid separation with high collection efficiency of proteins from cell lysates. This method was used to study the differences in protein profiles among normal cells and fully malignant cells that share a common genetic background. The highly expressed proteins in each cell type were separated and collected in the liquid state where they were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) to obtain the molecular weight of the proteins. The protein fractions were then subjected to tryptic digestion and analyzed by pulsed delay extraction (PDE)-MALDI-TOFMS to obtain the peptide maps. The expressed proteins were identified based upon the molecular weight and peptide map using database-searching procedures. It is shown that key cancer-related proteins can be detected and identified which may be potentially used as biomarkers for cancer detection.

Biomarkers, Tumor↗

PubMed1983

Morphological characteristics and protein profile of isolated human decidual cells.

Isolated decidual cells were prepared from human decidual tissue obtained during early pregnancy by digesting the tissue fragments with 0.1% collagenase solution. Morphological studies of the cells were carried out using morphometric and flow cytometric analysis while the protein profile was analysed by polyacrylamide gel electrophoresis and gel filtration column chromatography. An average of 90% cell viability was achieved and the results showed that decidual cells constitute up to 70% of cell number and 89% of cell area of the isolated decidual cell suspension. The presence of serum proteins in uterine tissue extracts is due to blood contamination. However, the similarities of the non-serum protein profiles in tissue and cellular extracts validates previous studies performed in uterine fluids and tissue extracts. Finally, at least one unique uterine protein appeared to be a sub-unit of a larger molecule.

Cell Count↗

PubMed2004

Quantitative protein profiling using antibody arrays.

Traditional approaches to microarrays rely on direct binding assays where the extent of hybridisation and the signal detected are a measure of the analyte concentration in the experimental sample. This approach, directly imported from the nucleic acid field, may fail if applied to antibody-antigen interactions due to the shortage of characterised antibodies, the significant heterogeneity of antibody affinities, their dependence on the extent of protein modification during labelling and the inherent antibody cross-reactivity. These problems can potentially limit the multiplexing capabilities of protein affinity assays and in many cases rule out quantitative protein profiling using antibody microarrays. A number of approaches aimed at achieving quantitative protein profiling in a multiplex format have been reported recently. Of those reported, the three most promising routes include signal amplification, multicolour detection and competitive displacement approaches to multiplex affinity assays. One in particular, competitive displacement, also overcomes the problems associated with quantitation of affinity interactions and provides the most generic approach to highly parallel affinity assays, including antibody arrays.

Animals↗