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PubMed1992

Changes in the protein profile and antigenicity of different Borrelia burgdorferi strains after reintroduction to Ixodes ricinus ticks.

Eight Swiss strains of Borrelia burgdorferi, with various protein profiles and the North-American strain B31 were artificially introduced into Ixodes ricinus ticks and reisolated 10 days later. All isolates were subsequently examined by SDS-PAGE analysis. Comparing initial isolates with the reisolates, we observed that 7 out of 9 strains changed their protein pattern with respect to the major proteins OspA, OspB and the 22 kDa protein after passage in the tick. The strains NE2, NE4 and NE83 with the initial phenotype of OspA and 22 kDa proteins changed to the phenotype of OspA and OspB, the strains B2 and NE202 with the initial phenotype of OspA acquired an additional protein of 22 kDa and the strain NE58 with the initial phenotype of OspA also acquired a protein of 22 kDa. Examination of these isolates by Western blot analysis demonstrated that the reaction with the monoclonal antibody H5332 and a monospecific polyclonal antibody PoAb/anti-22 kDa differed between the initial isolates and the reisolates.

Animals↗

PubMed1998

Protein profiles and antifungal activities of kernel extracts from corn genotypes resistant and susceptible to Aspergillus flavus.

Mechanisms of resistance to infection by the fungus Aspergillus flavus and accumulation of aflatoxin were studied in kernels of resistant (GT-MAS:gk, Mp420) and susceptible ( Pioneer 3154, Deltapine G-4666) corn genotypes. Proteins from kernel extracts of corn genotypes were analyzed by several methods of polyacrylamide gel electrophoresis. Consistent differences in protein profiles were detected among genotypes. Several proteins were unique to or present in greater concentration in resistant genotypes, whereas others were present only in susceptible genotypes. Extracts of resistant kernels showed markedly greater antifungal activity against A. flavus than did susceptible kernel extracts. Results from the present study suggest a role for kernel proteins in resistance to A. flavus infection and aflatoxin contamination in corn genotypes.

Aspergillus flavus↗

PubMed1990

Outer membrane protein profiles of Edwardsiella ictaluri from fish.

Outer membrane proteins (OMP) prepared with sodium N-lauroyl sarcocinate (SLS) from 33 Edwardsiella ictaluri isolates from fish were examined by electrophoresis. Twenty-eight isolates from channel catfish (Ictalurus punctatus) had similar OMP profiles. Ten bands (71 kilodaltons [kD] to 19.5 kD) were identified in all isolates from channel catfish. One major 35-kD protein comprised most of the protein content of the outer membrane of isolates from channel catfish. Differences existed among isolates in the amount of protein within minor OMP bands. Edwardsiella ictaluri ATCC 33202 contained larger quantities of the 38.5- and 37-kD proteins than did the other isolates. Outer membrane protein profiles of E ictaluri derived from Bengal danio (Danio devario) and walking catfish (Clarias batrachus) were identical to OMP profiles of isolates from channel catfish. In contrast, OMP profiles from single isolates from green knife fish (Eigemannia virescens) and white catfish (Ictalurus catus) were different. Variations in incubation time, SLS extraction time, SLS extraction number, and in vivo and in vitro passage had no effect on the OMP profile of E ictaluri ATCC 33202. An increase in duration of sample solubilization did affect the OMP profile of E ictaluri ATCC 33202 by decreasing the amount of protein in 52-, 46-, and 43.5-kD bands. Accompanying the decrease were increased staining intensity in the 31.5- and 28.5-kD bands and the appearance of 4 new bands (34, 33, 25.5, and 22.5 kD). Edwardsiella ictaluri, a gram-negative bacterium in the family Enterobacteriaceae, is the cause of enteric septicemia of catfish.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1986

Quantitative protein profiling: determining lexotypes.

The lexotype of a cell is defined as a set of quantitative characters of its informational macromolecular gene products, notably proteins, as observed under specified environmental conditions. This definition can be applied to cells in several ways that need to be distinguished. It can refer to the protein lexotype, to RNA lexotypes; to the steady-state lexotype, synthesis lexotype, functional protein lexotype; to the in situ lexotype and standard-environment lexotype. When used without qualification, the term lexotype may be applied to the standard-environment, steady-state protein lexotype. Some difficulties that currently limit our ability to determine lexotypes are assessed. Reasons are given why abnormal cellular states, such as states of disease, should often be characterizable by means of protein markers not themselves involved in the disease process and why one expects to find markers in tissues other than the one in which a certain pathological process may be anticipated to occur. There are three routes through which biological systems can produce secondary protein markers, namely through gene regulatory chains, through chromosomal gene linkage, and through "physiological linkage" of genes. The partly stable, partly shifting, yet defined relations between tissue lexotypes are considered. A number of potentially important fields of application of rigorous quantitative analyses of protein profiles are listed. One particular use of the technology is to investigate a hypothesis linking aging to degenerative diseases with late onset. According to this hypothesis, such diseases appear in later life as the cellular concentration of the active form of a protein passes a certain threshold in the course of the aging process.

Aging↗

PubMed1991

Organ and species specific differences in cytoskeletal protein profiles of cultured microvascular endothelial cells.

1. Using two-dimensional gel electrophoresis and immunoblotting techniques we systematically document the structural diversity of cytoskeletal proteins in tight and leaky cultured microvascular endothelial cells (MEC). Bovine pulmonary and eel rete mirabile MEC primarily express cytokeratins 8 and 19. Cytokeratins 8 and 18 were found to be prominent in rat pulmonary MEC. Bovine retinal MEC contained cytokeratins 8, 18 and 19. Bovine adrenal MEC contain vimentin as their sole intermediate filament protein. 2. Four principal actin isoforms were resolved in micro/macrovascular endothelial cells as well as in vascular smooth muscle cells. Retinal pericytes expressed three principal actin isoforms. 3. These results indicate that MEC are diverse, highly differentiated cells displaying a large repertoire of cytoskeletal protein profiles suited for specific tissue functions.

Actins↗

PubMed1976

Plasma protein profiling: the diagnostic evaluation of disorders in plasma protein composition by a new immunoelectrophoretic method.

A new electroimmunoprecipitation technique is presented by means of which a great variety of antigens e.g. plasma proteins can be simultaneously and quantitatively determined with a single-step electrophoretic separation. The essential features of the new technique are: (a) subdivision of the antibody gel into gel strips containing monospecific antibodies to individual plasma proteins. (b) sample application as a "sample gel" filling a trough over the width of the immunoplate. Quantitation is based on the fact that the distance an antigen can migrate within a gel containing a defined amount of specific antibody directed against the antigen is determined by the concentration of the appropriate antigen within the sample. The area where the antigen is finally completely consumed by immunoprecipitation and antibody present in excess is sharply delineated. The applicability of the method in simultaneous quantitative determination of 15 plasma proteins is demonstrated with plasma from healthy blood donors and patients with various diseases. The advantage of the new technique as compared to commonly used clinical acetate folia electrophoresis is the high degree of specificity for the determination of a great number of individual, diagnostically meaningful plasma proteins. The advantage over common quantitative two-dimensional immunoelectrophoresis is its uncomplicated way of evaluation. The potential clinical application of the new quantitative immunoelectrophoretic technique in diagnostic screening and differential diagnosis is discussed.

Blood Proteins↗

PubMedDate not supplied

A rapid protein profiling system that speeds study of cancer and other diseases.

To realize its potential to transform research and medicine, genomics must be complemented by protein expression data. Analysis by SELDI-TOF-MS technology, implemented in the ProteinChip System, overcomes the limitations of traditional protein analysis methods by enabling rapid analysis of small amounts of crude protein extracts and providing size and functional characterization on total cellular or tissue populations of proteins. The system has already been put to constructive use in many laboratories and has enabled researchers to determine unique protein profile signatures characteristic of progression from healthy to diseased states and back. In addition to its research applications, recent experimental successes suggest that the technology may soon find use in the important work of early diagnosis and treatment of infectious diseases, cancer, and other medical conditions.

Antibodies↗

PubMedDate not supplied

Sperm surface protein profiles of fertile and infertile men: search for a diagnostic molecular marker.

The present study was conducted to examine whether there is any difference in the protein profiles of sperm membranes of fertile (n = 37) and infertile (n = 29) men. In the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) procedure, the sperm extracts of fertile and infertile men showed 12-16 major and 8-14 minor common protein bands in various molecular regions. However, there was an additional band of approximately 78 kD present in the sperm extracts of approximately 80% of infertile men that was absent in the sperm extracts of fertile men. In the remaining approximately 20% of the infertile men, it was of relatively weaker intensity. This band was present only in the sperm extracts and not in the seminal plasma, whether of fertile or infertile men, and was designated as the infertility-associated sperm protein (IASP). Antibodies to IASP specifically immunoprecipitated the approximately 78-kD protein from the sperm extracts of only infertile and not the fertile men. In the immunofluorescence technique, the anti-IASP antibodies reacted strongly with the head and/or midpiece regions of morphologically abnormal sperm and cross-reacted with a protein in the equatorial regions of morphologically normal sperm of fertile men. The IASP molecule and its antibodies may provide molecular tools to differentiate between the normal and abnormal sperm, and, thus, may be useful markers for the diagnosis of male-factor infertility.

Biomarkers↗

PubMed1992

Analysis of direct tissue isoelectric focused protein profiles of resected intestinal mucosa and endoscopic biopsies from patients with inflammatory bowel disease.

Direct tissue isoelectric focusing was used as a procedure to analyze differences in soluble tissue protein profiles of resected intestinal segments and endoscopic biopsies from patients with ulcerative colitis, Crohn's disease, and colonic cancer. Extraction of tissue proteins was accomplished by electrophoresis of mucosal cryostat sections on agarose gels across a broad pH gradient. The inflamed colonic mucosa from Crohn's disease patients showed similar isoelectric focusing protein patterns. Small bowel mucosa from a patient with both colonic diverticular disease and Crohn's disease showed protein patterns identical with that of the mucosa from a patient with only Crohn's disease. The inflamed mucosae from ulcerative colitis patients revealed identical protein patterns but were distinct from those of non-inflamed ulcerative colitis mucosa and from the inflamed mucosae from Crohn's disease patients. Non-inflamed small bowel mucosae from cancer, ulcerative colitis, and Crohn's disease patients showed distinct protein patterns which were absent in the non-inflamed large bowel mucosae. The inflamed resected ileum of a Crohn's disease patient exhibited protein patterns similar to those of the biopsy of an inflamed mid-transverse large bowel. Mucosal biopsies from inflamed sigmoid colon of a Crohn's disease patient showed different protein patterns than those in biopsies from the inflamed mid-transverse colon. Thus, distinctive isoelectric focusing protein patterns may be useful in differentiating Crohn's colitis and ulcerative colitis when granulomata are absent, and in resolving indeterminant colitis to one of these classic inflammatory bowel diseases.

Biopsy↗

PubMed1986

Protein profiles of sarcoplasmic reticulum from normal and dystrophic mouse muscle.

Sarcoplasmic reticulum (SR) was isolated from skeletal muscle of dystrophic (C57BL/6J dy2J/dy2J) mice and the protein composition analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Densitometric analysis of dystrophic SR preparations indicated a decrease in the Ca2+-ATPase and calsequestrin, and the appearance of a protein with molecular weight 72 000. These differences in the protein profiles between normal and dystrophic SR became more apparent as the disease progressed. The observations are discussed in relation to secondary changes in the dystrophic process such as changes in fibre type and the presence of immature fibres.

Animals↗

PubMed2005

Evaluation of two-dimensional electrophoresis and liquid chromatography--tandem mass spectrometry for tissue-specific protein profiling of laser-microdissected plant samples.

Laser microdissection (LM) allows the collection of homogeneous tissue- and cell-specific plant samples. The employment of this technique with subsequent protein analysis has thus far not been reported for plant tissues, probably due to the difficulties associated with defining a reasonable cellular morphology and, in parallel, allowing efficient protein extraction from tissue samples. The relatively large sample amount needed for successful proteome analysis is an additional issue that complicates protein profiling on a tissue- or even cell-specific level. In contrast to transcript profiling that can be performed from very small sample amounts due to efficient amplification strategies, there is as yet no amplification procedure for proteins available. In the current study, we compared different tissue preparation techniques prior to LM/laser pressure catapulting (LMPC) with respect to their suitability for protein retrieval. Cryo-sectioning was identified as the best compromise between tissue morphology and effective protein extraction. After collection of vascular bundles from Arabidopsis thaliana stem tissue by LMPC, proteins were extracted and subjected to protein analysis, either by classical two-dimensional gel electrophoresis (2-DE), or by high-efficiency liquid chromatography (LC) in conjunction with tandem mass spectrometry (MS/MS). Our results demonstrate that both methods can be used with LMPC collected plant material. But because of the significantly lower sample amount required for LC-MS/MS than for 2-DE, the combination of LMPC and LC-MS/MS has a higher potential to promote comprehensive proteome analysis of specific plant tissues.

Arabidopsis↗

PubMed1977

[The regulation of acute inflammatory processes by systemic changes of plasma protein profiles].

Local inflammatory processes in the human as well as in experimental animals cause a selective increase or decrease of the hepatic synthesis rate of many plasma proteins. The resulting systemic changes in the plasma-protein profile regulate the extent of the local inflammatory response. The physiologic importance of this feedback mechanism is directed at preventing the spread of local inflammatory tissue destruction. By means of mediator substances involved in this regulatory system, the infalmmatory responses can be experimentally manipulated.

Animals↗

PubMed2003

Activity-based protein profiling in vivo using a copper(i)-catalyzed azide-alkyne [3 + 2] cycloaddition.

Toward the goal of assigning function to the tens of thousands of protein products encoded by eukaryotic and prokaryotic genomes, the field of proteomics requires new technologies that can functionally characterize proteins within the dynamic environment of the cell, where these biomolecules are subject to myriad posttranslational modifications and the actions of endogenous activators and inhibitors. Here, we report an advanced strategy for activity-based protein profiling (ABPP) that addresses this important need. We show that several enzymes can be labeled in an activity-based manner both in vitro and in vivo by an azido-sulfonate ester probe and that these labeling events can be detected in whole proteomes by copper-catalyzed ligation with a rhodamine-alkyne reagent. This click chemistry-based strategy for ABPP represents a unique and versatile method for functional proteome analysis.

Aldehyde Dehydrogenase↗

PubMed1978

Twelve serum protein profiles in children with acute nonbacterial gastroenterocolitis.

In thirty children hospitalized with acute benign, short-duration gastroenterocolitis, no obligate pathogens were isolated from stools. Five bleedings were established from each patient in order to obtain the protein profiles of albumin, orosomucoid, haptoglobin, alpha2-macroglobulin, ceruloplasmin, transferrin, C3-component, C-reactive protein, immunglobulins IgG, IgA, IgM and IgD. The proteins were quantitated by the single radial immunodiffusion method. The initial drop in some of the proteins followed may be related to general protein loss, negative nitrogen balance or hemodilution. The absence of a significant increase in all the investigated immunoglobulin classes contrasted with remarkable increase in haptoglobin and orosomucoid, both reaching normal levels in late convalescence. C-reactive protein could be demonstrated in half of the children showing early normalization with disappearance of clinical symptoms. In contrast to ceruloplasmin and C3- component, alpha2-macroglobulin was not involved in the acute phase protein reaction.

Acute Disease↗

PubMed1995

The relationship between the pressor response to involuntary isometric exercise and the contractile protein profile of the active muscle in man.

The present investigation examined the relationship between the pressor response during electrically evoked isometric ankle plantar flexion and the contractile protein profile of the active muscle in seven young men [mean (SD) age, 26 (6) years] and five older men [70 (4) years]. Muscle biopsy samples were taken from lateral gastrocnemius (LG) and soleus (SOL) of each subject. These were analysed for isomyosin composition using non-denaturing pyrophosphate polyacrylamide gel electrophoresis. The degree of association was examined between the cardiovascular changes and the fast isomyosin content of LG and SOL individually and in combination (SOL/LG). In the total subject group there was no association between the heart rate response or the change in systolic blood pressure (BP) and the fast isomyosin composition. However, the change in diastolic BP was significantly associated with the fast isomyosin composition of SOL/LG (delta diastolicBP = 0.31 + 0.045% FM SOL/LG, r = 0.65, P = 0.029). These findings suggest that the magnitude of the peripheral reflex mediated pressor response to isometric exercise and the fast isomyosin content of the active muscle are related.

Adult↗

PubMed2003

[Comparison of protein profiles and antibiotic sensitivity of methicillin-resistant Staphylococcus aureus strains isolated from nasal cultures of polyclinic patients and hospital personnel].

In this study, the antibiotic susceptibilities and whole cell protein patterns of methicillin resistant Staphylococcus aureus (MRSA) strains which were isolated from nasal swabs of outpatients and healthy hospital staff, have been determined by disk diffusion test and sodium dodecyl sulfate polyacrylamide gel electrophoresis method, respectively. A total of 43 MRSA isolates of which 13 out of 500 outpatients and 30 out of 500 hospital personnel, showed the same protein profiles, and the antibiotic susceptibility patterns were found similar. These findings suggested that the strains isolated from the outpatients were originated from hospital environment.

Ambulatory Care↗

PubMed2006

Informatics-assisted protein profiling in a transgenic mouse model of amyotrophic lateral sclerosis.

One of the causes of amyotrophic lateral sclerosis (ALS) is due to mutations in Cu,Zn-superoxide dismutase (SOD1). The mutant protein exhibits a toxic gain of function that adversely affects the function of neurons in the spinal cord, brain stem, and motor cortex. A proteomic analysis of protein expression in a widely used mouse model of ALS was undertaken to identify differences in protein expression in the spinal cords of mice expressing a mutant protein with the G93A mutation found in human ALS. Protein profiling was done on soluble and particulate fractions of spinal cord extracts using high throughput two-dimensional liquid chromatography coupled to tandem mass spectrometry. An integrated proteomics-informatics platform was used to identify relevant differences in protein expression based upon the abundance of peptides identified by database searching of mass spectrometry data. Changes in the expression of proteins associated with mitochondria were particularly prevalent in spinal cord proteins from both mutant G93A-SOD1 and wild-type SOD1 transgenic mice. G93A-SOD1 mouse spinal cord also exhibited differences in proteins associated with metabolism, protein kinase regulation, antioxidant activity, and lysosomes. Using gene ontology analysis, we found an overlap of changes in mRNA expression in presymptomatic mice (from microarray analysis) in three different gene categories. These included selected protein kinase signaling systems, ATP-driven ion transport, and neurotransmission. Therefore, alterations in selected cellular processes are detectable before symptomatic onset in ALS mouse models. However, in late stage disease, mRNA expression analysis did not reveal significant changes in mitochondrial gene expression but did reveal concordant changes in lipid metabolism, lysosomes, and the regulation of neurotransmission. Thus, concordance of proteomic and mRNA expression data within multiple categories validates the use of gene ontology analysis to compare different types of "omic" data.

Amyotrophic Lateral Sclerosis↗

PubMed1992

Observations on the interleukin-6 and acute phase protein profiles in the disease course of patients with lupus erythematosus.

In vitro models have shown that interleukin-6 (IL-6) is the main dominator of the stimulation of the full spectrum of acute phase proteins. This study describes IL-6 levels in relation to levels of acute phase proteins in 15 systemic lupus erythematosus (SLE) patients, with special attention given to those patients with increased serum levels of IL-6. Three episodes with elevated levels of IL-6 were observed in a period shortly after a flare-up of SLE, in three of the 15 patients. In one of these three patients a clear increase in the C-reactive protein (CRP) level, preceded by an IL-6 increase, was observed. In the other two patients, CRP levels remained unchanged. It is speculated that, next to IL-6, another signal is operative or needed for the start of an acute phase reaction. However, influences of the disease itself or of the administered therapy cannot be excluded as the cause of the described discrepancy between IL-6 and acute phase protein profiles in these two SLE patients.

Acute-Phase Proteins↗