Extraction and Partial Characterization of Lectin from Indonesian Brown Algae Padina australis and Padina minor

Extraction and partial characterization of lectin from Indonesian Padina australis and Padina minor had been carried out. The crude extract of the P. australis and P. minor were examined for hemagglutination activity (HA) using native and trypsin-treated of rabbit and human A, B, O type erythrocytes. Both extracts agglutinated all of the trypsin-treated erythrocytes tested in the HA assay. Strong HA was detected in the crude extract of P. minor with trypsin-treated of human type A and O erythrocytes. However, the sugar-binding specificity study through the quantitative hemagglutination inhibition (HI) assay showed that P. minor extract could not specifically recognize the glycans tested. Apparently, the HA of the P. minor was more due to its co-extracted polyphenols content than its lectin content. On the other hand, the HI assay showed that asialo transferrin human (aTf) and asialo porcine thyroglobulin (aPTG) were the most powerful in inhibiting the HA of P. australis. Those indicated that P. australis protein extract was able to specifically recognized aTf and aPTG. The stability of P. australis and P. minor HA over various temperatures, pH ranges, and divalent cations studies showed that the P. minor HA was stable on a wide range of pH and temperature; not affected by the presence of EDTA, but decreased by Ca2+ and Mg2+ additions showed that P. minor protein extract was not a metallic protein. The HA of P. australis decreased at 60 oC and was inactivated at 90 oC; increased at strong acidic (pH 3 & 4) and strong basic (pH 9 & 10) and dependent by the presence of either EDTA or Ca2+ and Mg2+ divalent cation.

Production of Recombinant EAV with Tagged Structural Protein Gp3 to Study Artervirus Minor Protein Localization in Infected Cells

Equine arteritis virus (EAV) is a prototype member of the Arterivirus family, comprising important pathogens of domestic animals. Minor glycoproteins of Arteriviruses are responsible for virus entry and cellular tropism. The experimental methods for studying minor Arterivirus proteins are limited because of the lack of antibodies and nested open reading frames (ORFs). In this study, we generated recombinant EAV with separated ORFs 3 and 4, and Gp3 carrying HA-tag (Gp3-HA). The recombinant viruses were stable on passaging and replicated in titers similar to the wild-type EAV. Gp3-HA was incorporated into the virion particles as monomers and as a Gp2/Gp3-HA/Gp4 trimer. Gp3-HA localized in ER and, to a lesser extent, in the Golgi, it also co-localized with the E protein but not with the N protein. The co-localization of Gp3-HA and the E protein with ERGIC was reduced. Moreover, EAV with Gp3-HA could become a valuable research tool for identifying host cell factors during infection and the role of Gp3 in virus attachment and entry.

Profil Protein Lima Jenis Daging yang Direndam Daun Pepaya Berbasis SDS-PAGE

Meat is an important food for fulfill nutritional needs, many of meats are consumed as a source of highest quality nutrition for humans, especially as a source of protein. Papaya leaves contain the enzyme papain (a protase enzyme that can hydrolyze proteins), so that it can be used to soften meat. The purpose of this study was to look at an overview of protein profiles in five types of meat, namely goat, beef, buffalo, free-range chicken and broiler chicken which were soaked in papaya leaves. The protein profile of five types of meat was analyzed using the SDS-PAGE 12% method. The results showed that the control meat of goat, beef, buffalo, free-range chicken and broiler chicken which were not soaked in papaya leaves showed that there were many major protein bands compared to minor protein bands. Whereas in goat, beef, buffalo, free-range chicken and broiler chicken which have been soaked in papaya leaves, the results were different compared to the control, there were many minor protein bands. While the major bands only have 6 to 9 protein bands. Based on these results indicate that immersion with the enzyme papain contained in papaya leaves can break down peptide bonds, if it works on meat it can be broken down so the meat becomes tender and protein bands in the form of micromolecules.Daging merupakan bahan pangan yang penting dalam memenuhi kebutuhan gizi, banyak dikomsumsi sebagai sumber nutrisi yang berkualitas bagi manusia terutama sebagai sumber protein. Daun pepaya mengandung enzim papain (enzim protase yang dapat menghidrolisa protein), sehingga dapat digunakan untuk melunakkan daging. Tujuan penelitian ini untuk melihat gambaran profil protein pada lima jenis daging yaitu daging kambing, sapi, kerbau, ayam kampung dan ayam potong yang direndam daun pepaya. Profil protein lima macam daging dianalisis menggunakan metode SDS-PAGE 12%. Hasil penelitian menunjukkan pada daging kontrol yaitu daging kambing, sapi, kerbau, ayam kampung dan ayam potong yang tidak direndam daun pepaya menunjukkan terdapat banyak pita protein mayor dibandingkan pita protein minor. Sedangkan pada daging kambing, sapi, kerbau, ayam kampung dan ayam potong yang telah direndam daun pepaya menunjukkan hasil yang berbeda dibandingkan dengan kontrol yaitu pada semua daging terdapat banyak pita protein minor. Sedangkan pita mayor hanya terdapat 6 sampai 9 pita protein saja. Berdasarkan hasil tersebut menunjukkan bahwa perendaman dengan enzim papain yang terdapat dalam daun pepaya dapat memecah ikatan peptida, jika bekerja pada daging dapat diuraikan sehingga daging menjadi empuk, dan pita protein berbentuk mikromolekul.

Sequence analysis of minor protein genes of equine arteritis virus during persistent infection

The variability of the ORF2a, ORF2b, ORF3, and ORF4 genes of the equine arteritis virus (EAV) was analysed during a seven year observation of persistent infection in a stallion of the Malopolska breed. A total of 11 semen samples were collected between 2004 and 2011. RNA of EAV isolates obtained from the semen of the stallion was amplified, sequenced, and compared with the sequences of other strains available in GenBank. Multiple nucleotide substitutions were found in sequences of the analysed regions, however, neither deletion nor insertions were detected. The highest number of point mutations (11-6 synonymous and 5 non-synonymous) were found in the ORF2b gene, and the lowest number of substitutions (6-5 synonymous and one non-synonymous) were found in the ORF2a gene. None of the identified mutations affected any of the glycosylation or phosphorylation sites of the minor EAV protein. Phylogenetic analysis of the ORF3 gene of EAV isolates showed that they grouped together within the cluster of European strains of EAV. Additionally, the ORF3 gene sequences of the isolates showed high (86.4% - 98.3%) similarity to the previously isolated Polish EAV strains.

Polyacrylamide gel electrophoresis of soybean seed proteins

Four major and 14 minor protein bands were detected when total salt soluble proteins of soybean (Glycine max cultivar Warszawska) seed were subjected to polyacrylamide gel electrophoresis under nondissociating conditions, and 16 protein bands were detected under dissociating conditions. Molecular weights of three major protein fractions in PAGE SDS were determined for around 18 500, 36 000 and 80 000 daltons.

Motility Is Crucial for the Infectious Life Cycle of Borrelia burgdorferi

ABSTRACTThe Lyme disease spirochete,Borrelia burgdorferi, exists in a zoonotic cycle involving an arthropod tick and mammalian host. Dissemination of the organism within and between these hosts depends upon the spirochete's ability to traverse through complex tissues. Additionally, the spirochete outruns the host immune cells while migrating through the dermis, suggesting the importance ofB. burgdorferimotility in evading host clearance.B. burgdorferi's periplasmic flagellar filaments are composed primarily of a major protein, FlaB, and minor protein, FlaA. By constructing aflaBmutant that is nonmotile, we investigated for the first time the absolute requirement for motility in the mouse-tick life cycle ofB. burgdorferi. We found that whereas wild-type cells are motile and have a flat-wave morphology, mutant cells were nonmotile and rod shaped. These mutants were unable to establish infection in C3H/HeN mice via either needle injection or tick bite. In addition, these mutants had decreased viability in fed ticks. Our studies provide substantial evidence that the periplasmic flagella, and consequently motility, are critical not only for optimal survival in ticks but also for infection of the mammalian host by the arthropod tick vector.

THE INFLUENCE OF ORGANIC SOLVENT PROTEIN PRECIPITATION ON SDS PAGE PROTEIN PROFILE IN SERUM

A study on the influence of organic solvent protein precipitation to the profile of the serum protein has been accomplished. The expected conditions were precipitation of abundant proteins present in serum result in increasing relative concentration of minor protein which can be useful for sample preparation for biomarker studies. The serum were precipitated with various diluted (