scholarly journals Comparative proteomic study of pig muscle proteins during growth and development of an animal

2022 ◽  
Vol 6 (4) ◽  
pp. 320-327
Author(s):  
A. G. Akhremko ◽  
E. S. Vetrova
Keyword(s):  
Muscle Tissue ◽  
Growth And Development ◽  
Mass Spectrometric ◽  
Protein Fractions ◽  
Two Dimensional ◽  
Weaned Pigs ◽  
Muscle Sample ◽  
One Dimensional ◽  
Significant Difference ◽  
Dimensional Electrophoresis

The production of high-quality pork is closely related to the growth and development of muscle tissue. The present article provides a comparative proteomic research of l. dorsi, b. femoris, m. brachiocephalicus during the pigs’ growth and development (at age of 60 days and 180 days). This work was supported by data of electrophoretic methods: one-dimensional electrophoresis according to Laemmli with densitometric assessment in the ImageJ software and two-dimensional electrophoresis according to O’Farrell method with its further processing on the software ImageMaster. The mass spectrometric identification was conducted with the help of the high-performance liquid chromatography (HPLC) system connected to a mass spectrometer; further the data were interpreted by search algorithm Andromeda. When comparing frequency diagrams of one-dimensional electrophoregrams of all three muscle tissues of weaned pigs, the greatest difference was observed for the muscle sample l. dorsi. Comparison of diagrams of muscle tissue samples taken for mature pigs showed a great similarity of all three studied muscles samples. Within the framework of the research, the Fold indicator was calculated. The exceeding its value by more than 2 units is generally considered to be a statistically significant difference. When analyzing two-dimensional electrophoretograms of weaned pigs’ muscles, 18 protein fractions were revealed with Fold > 2. When examining the muscle tissue of mature pigs, 15 of those proteins were found; the differences were mostly detected in the minor protein fractions. The mass spectrometric analysis of the cut bands with well-pronounced differences from the onedimensional electrophoretogram revealed 214 proteins involved to a greater extent in cellular and metabolic processes, physical activity and localization. Growth and development protein — semaphorin‑6B (96.78 kDa) — was revealed in muscle tissue of l. dorsi, a. Also in l. dorsi and b. femoris the growth and development proteins were found: cadherin‑13 (78.23 kDa), cadherin‑7 (87.01 kDa), the F‑actin-cap protein beta subunit (30.66 kDa), and two uncharacterized proteins at 65.60 kDa and 63.88 kDa.

scholarly journals Ultrastructural localization of giardins to the edges of disk microribbons of Giarida lamblia and the nucleotide and deduced protein sequence of alpha giardin.

1989 ◽  
Vol 109 (5) ◽  
pp. 2323-2335 ◽  
Author(s):  
D A Peattie ◽  
R A Alonso ◽  
A Hein ◽  
J P Caulfield
Keyword(s):  
Protein Sequence ◽  
Protein Identification ◽  
Stop Codon ◽  
In Vitro Translation ◽  
Two Dimensional ◽  
One Dimensional ◽  
Amino Terminal ◽  
Ventral Disk ◽  
Dimensional Electrophoresis

The giardins are a group of 29-38-kD proteins in the ventral disk of the protozoan parasite Giardia lamblia. The disk attaches the parasite to the host's intestinal epithelium and is composed of parallel, coiled microtubules that are adjacent to the ventral plasma membrane and from which processes called microribbons extend into the cytoplasm; the microribbons are connected by crossbridges. G. lamblia cytoskeletons, consisting of disks and attached flagella, were isolated and used to show that the 29-38-kD proteins separate into five bands by one-dimensional electrophoresis and into 23 species by two-dimensional analysis. Rabbit antibodies raised against a 33-kD protein band, purified by one-dimensional gel electrophoresis and shown to contain three proteins by two-dimensional electrophoresis, recognized 17 proteins by two-dimensional immunoblot analysis. By immunofluorescence these antibodies reacted with the ventral disk but not with the flagella in isolated cytoskeletons. Electron microscopy revealed that the anti-giardin antibodies bound to the edges of the microribbons but not to the microtubules, crossbridges, or other, nondisk structures. Antibodies to tubulin reacted with both the disk and flagella in isolated cytoskeletons but bound only to the microtubules in these structures. The amino-terminal sequence of the 33-kD immunogen was determined and used to construct a DNA oligomer, and the oligomer was used to isolate the alpha giardin gene. The gene was used to hybrid select RNA, and the in vitro translation product from this RNA was precipitated by the antibodies against the 33-kD immunogen. The gene sequence was a single open reading frame of 885 nucleotides that predicted a protein of 33.8 kD. The protein sequence is unique, having no significant homology to two other giardin sequences or to any sequences within the Protein Identification Resource. It is predicted to be 82% alpha helical. The downstream sequence of the gene indicates that the sequence AGT-PuAA is located six to nine nucleotides beyond the stop codon in all protein-encoding genes of G. lamblia that have been sequenced and reported to date.

Plasma apolipoprotein pattern in fish-eye disease examined by high-resolution two-dimensional electrophoresis.

1985 ◽  
Vol 31 (12) ◽  
pp. 2032-2035 ◽  
Author(s):  
T Marshall ◽  
K M Williams ◽  
L Holmquist ◽  
L A Carlson ◽  
O Vesterberg
Keyword(s):  
High Resolution ◽  
Plasma Proteins ◽  
Eye Disease ◽  
Two Dimensional ◽  
Protein Patterns ◽  
Two Dimensional Electrophoresis ◽  
Significant Difference ◽  
Fish Eye ◽  
Dimensional Electrophoresis ◽  
Plasma Apolipoprotein

Abstract We compared the plasma protein patterns of the two living patients suffering from fish-eye disease with those of appropriate controls, using high-resolution two-dimensional electrophoresis. Quantitative abnormalities were detected in plasma polypeptides corresponding to the isoforms of apolipoproteins A-I and A-II. The disease was characterized by normal concentrations of proapo A-I but dramatically subnormal concentrations of the other apo A-I isoforms and apo A-II. No significant difference was detected in the concentrations of other plasma proteins. These findings are discussed in relation to other apolipoprotein disorders.

scholarly journals Adaptation of two-dimensional electrophoresis for muscle tissue analysis

10.5219/1380 ◽  
2020 ◽  
Vol 14 ◽  
pp. 595-601
Author(s):  
Anastasiya Akhremko ◽  
Ekaterina Romanovna Vasilevskaya ◽  
Liliya Fedulova
Keyword(s):  
Muscle Tissue ◽  
Protein Separation ◽  
Molecular Mechanisms ◽  
Raw Material ◽  
Bromophenol Blue ◽  
Tissue Analysis ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Bottom Chamber ◽  
Dimensional Electrophoresis

It is important to understand the molecular mechanisms that take place in muscle tissues and to predict meat quality characteristics. One of the most popular methods is two-dimensional electrophoresis, which allows us to visualize, share and identify different molecules, including meat proteins. However, the standard conditions of this method are not universal for all types of raw material, so the authors suggest a new variation of two-dimensional electrophoresis for muscle tissue analysis. Samples were tested by the classical version of isoelectric focusing (cathode buffer in the top and anode buffer in the bottom chamber of the electrophoresis cell) and its variation (anode buffer in the top and cathode buffer in the bottom chamber of the electrophoresis cell). Next, extruded gels were incubated in two different buffer systems: the first was equilibration buffer I (6 M urea, 20% w/v glycerol, 2% w/v SDS and 1% w/v Ditiothreitol in 375 mM Tris-HCl buffer, pH 8.8) followed by equilibration buffer II (6 M urea, 20% w/v glycerol, 2% w/v SDS and 4% w/v iodoacetamide in 375 mM Tris-HCl buffer pH 8.8 and the second, buffer А, consisting of 5 M urea, 2% w/v SDS, 5% v/v mercaptoethanol, 62.5 mM Tris-HCl buffer, pH 6.8 and 0.01% w/v bromophenol blue. Electrophoretic studies of muscle tissue revealed the best protein separation after changing the direction of the current (authors' variation), while no differences were detected after changing incubation buffers.

scholarly journals Immunoglobulin G Subclasses and Prolactin (PRL) Isoforms in Macroprolactinemia Due to Anti-PRL Autoantibodies

2005 ◽  
Vol 90 (5) ◽  
pp. 3036-3044 ◽  
Author(s):  
Naoki Hattori ◽  
Katsuji Ikekubo ◽  
Yasuhisa Nakaya ◽  
Kaori Kitagawa ◽  
Chiyoko Inagaki
Keyword(s):  
Mass Spectrometry ◽  
Mass Spectrometric ◽  
Igg Subclasses ◽  
Two Dimensional ◽  
Human Pituitary ◽  
Two Dimensional Electrophoresis ◽  
Antigen Stimulation ◽  
Immunoglobulin G Subclasses ◽  
Specific Igg ◽  
Dimensional Electrophoresis

Although macroprolactinemia due to antiprolactin (anti-PRL) autoantibodies is not uncommon among hyperprolactinemic patients, the pathogenesis of such macroprolactinemia is still unknown. We examined IgG subclasses of anti-PRL autoantibodies by enzyme immunoassay, and PRL phosphorylation and isoforms by Western blotting, mass spectrometry, and two-dimensional electrophoresis in six patients with anti-PRL autoantibodies and in 29 controls. PRL-specific IgG subclasses in patients with anti-PRL autoantibodies were heterogeneous, but five of six patients showed IgG4 predominance, which is known to be produced by chronic antigen stimulation. Western blot and mass spectrometric analyses revealed that human pituitary PRL was phosphorylated at serine 194 and serine 163, whereas serine 163 in serum PRL was dephosphorylated. On two-dimensional electrophoresis, serum PRL mainly consisted of isoform with isoelectric point (pI) 6.58 in control hyperprolactinemic patients, whereas acidic isoforms (pIs 6.43 and 6.29) were also observed in patients with anti-PRL autoantibodies. Our data first demonstrate that human pituitary PRL is serine phosphorylated and partially dephosphorylated in serum, and suggest that the acidic isoforms may give rise to chronic antigen stimulation in patients with anti-PRL autoantibodies.

scholarly journals GENETIC VARIATION IN PROTEINS: COMPARISON OF ONE-DIMENSIONAL AND TWO-DIMENSIONAL GEL ELECTROPHORESIS

Genetics ◽  
1983 ◽  
Vol 104 (2) ◽  
pp. 381-390
Author(s):  
Tracy McLellan ◽  
Giovanna Ferro-Luzzi Ames ◽  
Kishiko Nikaido
Keyword(s):  
Genetic Variation ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
One Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Related Proteins ◽  
Dimensional Electrophoresis ◽  
Denaturation Of Proteins ◽  
New Mutations ◽  
Limited Sensitivity

ABSTRACT Two proteins with known characteristics on one-dimensional gels were studied by two-dimensional electrophoresis to compare the sensitivities of the two methods in detecting genetic variation. Two-dimensional electrophoresis was found to be less sensitive than several types of one-dimensional gels in distinguishing variants of both proteins. Denaturation of proteins in urea in the two-dimensional method makes it possible to distinguish closely related proteins that differ from each other by units of charge. Many more types of variation in protein sequences can be distinguished on one-dimensional gels in the absence of denaturants. The estimates of heterozygosity based on two-dimensional gels are lower than those based on other methods, at least in part, because of the limited types of sequence differences that can be detected on two-dimensional gels. The application of two-dimensional electrophoresis to the measurement of genetic variation and to the detection of new mutations should be made carefully, in view of the limited sensitivity of the method in finding differences in sequence.

scholarly journals Identification of mare colostrum proteins

2021 ◽  
Vol 19 (4) ◽  
pp. 25-32
Author(s):  
Weronika Medeńska ◽  
◽  
Alicja Dratwa-Chałupnik ◽  
Małgorzata Ożgo ◽  
Aleksandra Cichy ◽  
...  
Keyword(s):  
Growth And Development ◽  
Whey Proteins ◽  
Lactation Period ◽  
Gene Products ◽  
Two Dimensional ◽  
Immune Systems ◽  
Biochemical Processes ◽  
Two Dimensional Electrophoresis ◽  
Depth Analysis ◽  
Dimensional Electrophoresis

Colostrum is an essential feed of foals. It is a source of nutrients and functional proteins significant for foals’ growth and development. In the presented research using two-dimensional electrophoresis coupled via spectrometry mass MALDI-TOF in the mares’ colostrum (whey proteins fraction) were identified 24 proteins representing 15 different gene products. The identified proteins were involved in supporting foals’ immature immune systems and in the transport of various compounds. Further research of mares’ colostrum will allow determining more gene products. An in-depth analysis of mares’ milk will provide information about biochemical processes occurring in the mammary gland of the mare during the lactation period.

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