scholarly journals Prekeratin biosynthesis in human scalp epidermis

1982 ◽  
Vol 208 (1) ◽  
pp. 179-187 ◽  
Author(s):  
P T Bladon ◽  
P E Bowden ◽  
W J Cunliffe ◽  
E J Wood
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl ◽  
Post Translational Modification ◽  
Two Dimensional Gel Electrophoresis ◽  
Keratin Gene ◽  
One Dimensional ◽  
Electrophoretic Mobilities ◽  
Isoelectric Points ◽  
Intense Labelling ◽  
Dimensional Electrophoresis

Analysis of human scalp epidermal prekeratin polypeptides by two-dimensional gel electrophoresis revealed that each of the bands observed in one-dimensional electrophoresis consisted of three to five polypeptides of the same molecular weight but differing in isoelectric points. It was possible to divide the polypeptides into two families, with isoelectric points in the ranges pH 6.0-8.0 and pH 5.0-5.5 respectively. Incorporation of radiolabelled amino acids into freshly excised pieces of scalp epidermis showed that some of the polypeptides had relatively greater contents of glycine and serine than others. Radiolabelled methionine and leucine were, in contrast, incorporated more or less uniformly into all the polypeptides. After incubation with 32P-labelled orthophosphate, relatively more intense labelling by 32P was observed in the higher molecular weight bands of each family. The most basic of the isoelectric variants in each case did not take up phosphate, implying that at least some of the variation in charge was due to different degrees of phosphorylation. Polyadenylated RNA isolated from scalp epidermis was translated in an RNA-dependent reticulocyte haemolysate system followed by immunoprecipitation and electrophoresis. The polypeptides isolated by using anti-(human scalp prekeratin) immunoglobulin G had similar electrophoretic mobilities in sodium dodecyl sulphate/polyacrylamide gels to authentic prekeratin polypeptides, but had different isoelectric properties. This suggested that the products of keratin gene expression undergo post-translational modification.

scholarly journals Variations in the Methionine-rich Protein Composition of the Genus Arachis1

1984 ◽  
Vol 11 (1) ◽  
pp. 1-3 ◽  
Author(s):  
Sheikh M. Basha ◽  
Sunil K. Pancholy
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Protein Composition ◽  
Electrophoretic Analysis ◽  
Two Dimensional ◽  
Weight Group ◽  
Two Dimensional Gel Electrophoresis ◽  
One Dimensional ◽  
Molecular Weights ◽  
Isoelectric Points

Abstract Methionine-rich proteins (MRP) from seeds of different species of the Genus Arachis were isolated and analyzed by gel electrophoresis to detect possible compositional differences. One-dimensional gel electrophoretic analysis showed presence of quantitative and qualitative variations among the MRP-fractions. Following two-dimensional gel electrophoresis, the MRP-fractions were found to contain three groups of polypeptides with apparent molecular weights of approximately 21,000; 19,000 and 16,000, and isoelectric points between 5.1 and 5.8. Within each molecular weight group the number of polypeptides varied between 1 and 3.

PROTEIN PROFILING OF DIFFERENT PLANT TISSUES FROM HERB PHYLLANTHUS NIRURI

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Ainul Mardhiah Mohd Nail ◽  
Noor Hasniza Md Zin
Keyword(s):  
Molecular Weight ◽  
Protein Band ◽  
Protein Profiling ◽  
Protein Extract ◽  
Phyllanthus Niruri ◽  
Two Dimensional Gel Electrophoresis ◽  
Plant Parts ◽  
Sds Page ◽  
Isoelectric Points

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anticancer, antibacterial, antioxidant, anti-hypertensive and also anti-diabetic properties.  In this research, the proteomic part of P. niruri was studied to determine the bioactive peptides that responsible for specific characteristics. Total soluble proteins from different plant parts of freshly collected P. niruri were extracted using TCA/acetone method and then quantified using Bradford assay. Fruits part was found to have a significantly higher amount of proteins (4.91µg/µl + 0.21) compared to leaves (4.18µg/µl + 0.15). To determine the quality of proteins in the crude extract, SDS-Page was carried out which separates proteins in the basis of molecular weight. Proteins extracted from leaves were widely distributed between the range of 3.5 kDa to 160 kDA. Meanwhile, proteins in fruits mainly distributed within the range of 15 kDa to 80 kDa. The most highly expressed protein band was found in fruit, located in between 30 to 40 kDa. The protein extracts were then further analyzed based on the molecular weight and isoelectric points using two-dimensional gel electrophoresis (2D-GE) approach. Based on the profile pattern obtained from 2D-GE analysis, protein extract from fruits seems to express more protein spots compared to protein extract from leaves. Protein spots from fruit are seen to be intensely resolved within pH 4 to 10 at molecular weight between 10 kDa to 80 kDa. On the other hand, protein spots from leaves were moderately resolved at pH 4 to 10 at molecular weight within 10 kDa to 50 kDa.

Structure and chemical composition of insoluble filamentous components of sperm flagellar microtubules

1982 ◽  
Vol 58 (1) ◽  
pp. 1-22
Author(s):  
R.W. Linck ◽  
G.L. Langevin
Keyword(s):  
Peptide Mapping ◽  
Sodium Dodecyl ◽  
Protein Composition ◽  
Resistant Material ◽  
Remarkable Feature ◽  
Electrophoretic Mobilities ◽  
Helical Content ◽  
Isoelectric Points ◽  
Flagellar Proteins ◽  
Final Fraction

By progressive solvent extraction, we have obtained a series of subfragments of flagellar microtubules. Mild treatment gives rise to ribbons that contain longitudinally arranged protofilaments. Further extraction leaves a distinctive residue containing thinner ribbons, of three and eventually two protofilaments. Finally, filaments 2–3 nm in diameter and fibrous ribbons apparently containing 6 or more 2 nm subfibrils are found. This latter solvent-resistant material is consistently enriched in a characteristic set of polypeptides, which are found in flagella of several different species, including echinoderms and a mollusc. These polypeptides appear different from alpha- and beta-tubulin on the basis of their solubilities, isoelectric points and electrophoretic mobilities in sodium dodecyl sulphate/polyacrylamide gels; these conclusions are reinforced by peptide mapping after limited proteolytic digestion, although the latter method reveals certain similarities between these unique flagellar proteins, tubulin, chicken gizzard desmin and rabbit actin. A remarkable feature of the protein in the final fraction is the high alpha-helical content: 71% as measured by circular dichroism. We consider the possible origins of these filaments in the microtubule, in particular the possibility that microtubule protofilaments are heterogeneous in protein composition, and we discuss some of the implications of our findings.

Differences in Transcription and Translation of Long and Short GSα, the Stimulatory G-Protein, in Human Atrium

1995 ◽  
Vol 89 (5) ◽  
pp. 487-495 ◽  
Author(s):  
M. Susan Monteith ◽  
Tao Wang ◽  
Morris J. Brown
Keyword(s):  
Cyclic Amp ◽  
Isoelectric Point ◽  
Right Atrial ◽  
Two Dimensional ◽  
Human Atrium ◽  
Post Translational Modification ◽  
Two Dimensional Gel Electrophoresis ◽  
Significant Difference ◽  
Isoelectric Points

1. We have previously reported the relative mRNA and protein level of the long and short splice variants of Gsα (GsαL and GsαS) in human atrium. We have now measured the relative proportions of the serine+ and serine− variants of GsαL and GsαS in human atrium, and assessed, indirectly, whether their differential expression may (i) regulate Gsα phosphorylation, and (ii) be regulated by atrial cyclic AMP levels. 2. The serine+ and serine− variants of GsαL and GsαS were estimated by single nucleotide primer extension in 36 right atrial strips of which half were from β-adrenoceptor-blocked patients. The ratio of serine+ to serine− variants was 0.06 ± 0.12 for GsαL, compared with 8.04 ± 12.16 for GsαS (P < 0.001). 3. Isoelectric points of GsαL and GsαS in the atria of four β-adrenoceptor-blocked and four non-β-adrenoceptor-blocked patients were estimated by two-dimensional gel electrophoresis. Two-dimensional gel analysis gave a consistent pattern with several spots for both GsαL and GsαS; however, the isoelectric points of GsαS were more acid (5.18 ± 0.24) than those of GsαL (5.87 ± 0.17, P < 0.001). 4. No significant difference in either the serine variants or isoelectric point value was observed between β-adrenoceptor-blocked and non-β-adrenoceptor-blocked patients. 5. In conclusion, all four Gsα variants were expressed in human atrium, but GsαL is almost entirely of the serine− form. GsαS has a more acidic isoelectric point than GsαL, indicating a possible post-translational modification. The lack of difference in our results between β-adrenoceptor-blocked and non-β-adrenoceptor-blocked patients suggests indirectly that cyclic AMP is an unlikely candidate for regulating splicing or post-translational modification of Gsα in vivo.

The oral apparatus of Tetrahymena. V. Oral apparatus polypeptides and their distribution

1980 ◽  
Vol 44 (1) ◽  
pp. 317-333
Author(s):  
R.H. Gavin
Keyword(s):  
Molecular Weight ◽  
Basal Body ◽  
Tetrahymena Thermophila ◽  
Electrophoretic Analysis ◽  
Two Dimensional ◽  
Molecular Weight Range ◽  
Two Dimensional Electrophoresis ◽  
Weight Range ◽  
Isoelectric Points ◽  
Dimensional Electrophoresis

Two-dimensional electrophoresis was used to resolve approximately 162 polypeptides from the isolated oral apparatus of Tetrahymena thermophila. The molecular weight range was between 110 000 and 15 000 Daltons. The polypeptides had apparent isoelectric points between pH 3.3 and pH 7.2. Electrophoretic analysis of isolated ciliary axonemes and fractionated oral apparatuses made possible the assignment of polypeptides to structures within the oral apparatus. Approximately 24 polypeptides, including alpha and beta tubulins, are probable components of the basal body-basal plate complex. At least 5 of the oral apparatus polypeptides, including alpha and beta tubulin, are components of the oral apparatus ciliary axonemes. Approximately 138 polypeptides are components of the oral apparatus framework.

scholarly journals Identification of two major proteins of bovine pancreatic stones as immunoreactive forms of trypsinogens

1982 ◽  
Vol 205 (3) ◽  
pp. 543-549 ◽  
Author(s):  
A De Caro ◽  
L Multigner ◽  
H Vérine
Keyword(s):  
Molecular Weight ◽  
Isoelectric Focusing ◽  
Sodium Citrate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Charge Heterogeneity ◽  
Two Dimensional Electrophoresis ◽  
Pancreatic Stones ◽  
Dimensional Electrophoresis

Two major proteins have been identified in sodium citrate extracts of bovine pancreatic stones from 15 glands with lithiasis. They were found to have a molecular weight of about 24 000 and were further characterized by a variety of methods, including polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate, isoelectric focusing, two-dimensional electrophoresis, immunodiffusion, immunoelectrophoresis and determination of N-terminal residues. These two immunologically and electrophoretically different proteins were definitely shown to be immunoreactive forms of anionic and cationic trypsinogens, which are normal components of pancreatic juice. However, in contrast with both secretory trypsinogens, the stone proteins displayed an important charge heterogeneity under isoelectric-focusing conditions. A possible role for both secretory trypsinogens in pancreatic lithogenesis is suggested by the reproducibility of the data. Finally, two minor proteins with a lower molecular weight (about 11 000-13 000) have also been found to be present in all extracts, but have not yet been identified.

Isoelectric points and molecular weights of salt-extractable ribosomal proteins

1976 ◽  
Vol 54 (12) ◽  
pp. 1029-1033 ◽  
Author(s):  
M Saleem ◽  
Burr Atkinson
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Molecular Weights ◽  
Isoelectric Points ◽  
Acidic Proteins ◽  
Gel Isoelectric Focusing ◽  
Liver Ribosomes

Rat liver ribosomes prepared in low salt buffer contain basic and acidic proteins not found on ribosomes washed in high salt buffer. Proteins extracted from liver ribosomes by 500 mM KCl were characterized by acid urea–polyacrylamide gel electrophoresis, sodium dodecyl sulfate – polyacrylamide gel electrophoresis and gel isoelectric focusing. The salt-solubilized proteins contain 12 polypeptides with a molecular weight over 67 000, several polypeptides with molecular weights less than 67 000, and three polypeptides whose molecular weight exceeded 130 000. Ten to 12 of the proteins were basic, and about 24 acidic proteins were partially or wholly extracted from the ribosomes. Four of the acidic proteins have isoelectric points less than 4.5.

Proteins of human urine. II. Identification by two-dimensional electrophoresis of a new candidate marker for prostatic cancer.

1982 ◽  
Vol 28 (1) ◽  
pp. 160-163 ◽  
Author(s):  
J J Edwards ◽  
N G Anderson ◽  
S L Tollaksen ◽  
A C von Eschenbach ◽  
J Guevara
Keyword(s):  
Sodium Dodecyl ◽  
Prostatic Hyperplasia ◽  
Prostatic Adenocarcinoma ◽  
Relative Molecular Mass ◽  
Urogenital System ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Candidate Marker ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis

Abstract A protein series common to the urine and prostatic tissue of 16 of 17 patients with prostatic adenocarcinoma has been identified by high-resolution two-dimensional gel electrophoresis. These proteins, designated PCA-1, have a relative molecular mass in sodium dodecyl sulfate of about 40000. Analyses of urines from eight age-matched controls, seven patients with other ty pes of urogenital malignancies, two patients with benign prostatic hyperplasia, and five patients with malignancies not associated with the urogenital system failed to show PCA-1 in the patterns. These preliminary findings suggest that this protein should be systematically investigated as a candidate marker for prostatic adenocarcinoma in man.

scholarly journals Lyt-2 glycoprotein is synthesized as a single molecular species.

1983 ◽  
Vol 157 (1) ◽  
pp. 365-370 ◽  
Author(s):  
E Rothenberg ◽  
D Triglia
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Cell Surface ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Two Dimensional ◽  
Post Translational Modification ◽  
Molecular Weights ◽  
Gradient Electrophoresis ◽  
Isoelectric Points

We investigated the possibility that the Lyt-2 molecules made by uncloned mouse T lymphocytes would show variable primary structures like those of immunoglobulins. Newly synthesized Lyt-2/3 complexes were found to include only two major components, both discrete glycoproteins with apparent molecular weights of 31,000 (31 K) and 35,000 (35 K). When products of Lyt-2.1 and Lyt-2.2 thymocytes were compared by two-dimensional nonequilibrium pH gradient electrophoresis and sodium dodecyl sulfate polyacrylamide gel electrophoresis, the isoelectric points of the 35 K molecules were different; thus, the 35 K component was likely to be encoded by the Lyt-2 locus itself. However, the 35 K molecules made by any one genotype were homogeneous in charge as well as in size. The homogeneity was obscured rapidly by post-translational modification. Most strikingly, within 30 min of initial synthesis, these processing events generated the conspicuous array of microheterogeneous products that form the "38 K" component of cell-surface Lyt-2/3.

scholarly journals Polypeptide Composition of Arachin and Non-arachin Proteins From Early Bunch Peanut (Arachis hypogaea L.) Seed1

1981 ◽  
Vol 8 (2) ◽  
pp. 82-88 ◽  
Author(s):  
Shaik-M. M. Basha ◽  
Sunil K. Pancholy
Keyword(s):  
Molecular Weight ◽  
Arachis Hypogaea ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Seed Proteins ◽  
Two Dimensional ◽  
Polypeptide Composition ◽  
Two Dimensional Gel Electrophoresis ◽  
Arachis Hypogaea L ◽  
Isoelectric Points

Abstract Peanut (Arachis hypogaea L.) seed proteins were resolved into arachin and non-arachin fractions, and composite two-dimensional polypeptide maps were prepared. Seed proteins were extracted with a buffer containing 2 M NaCl, 10 mM Tris-HCl (pH 8.2), 0.2 mM phenylmethyl sulfonyl fluoride and 0.002% NaN3 and resolved into ten peaks by gel filtration on a Sephacryl S-300 column. Gel filtration of total protein extract yielded three molecular weight variants (490,000., 400,000, and 365,000) of arachin. Gel electrophoresis showed quantitative and qualitative differences in the protein and polypeptide composition of the three arachin variants. Nonarachin proteins obtained by this method were heterogeneous and distinct from the arachin. Two-dimensional gel electrophoresis revealed several differences in the polypeptide composition between arachin fraction IV and fractions II and III. Composite two-dimensional polypeptide maps of arachin and non-arachin revealed the presence of several polypeptides with similar isoelectric points and molecular weights between them. Arachin contained six molecular weight (between 15,500 and 68,000) classes of polypeptides with isoelectric points between 4.7 and 8.4 while nonarachin contained nine molecular weight (between 16,000 and 170,000) classes of polypeptides having isoelectric points between 4.7 and 7.9.

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