Masker Peel-Off Kulit Buah Jeruk Purut (Citrus hystrix) Sebagai Antiacne

Kulit buah jeruk purut (Cytrus Hystrix) biasa digunakan secara komersil sebagai perasa dan pengaroma karena minyak esensial memiliki kandungan hidrokarbon monoterpen, dengan komponen utama adalah β‐pinene (18.76%) dan limonene (30.73%) serta komponen lain yaitu terpinolene (4.33%), α‐terpinene (5.09%), γ‐terpinene (6.18%), terpineol (8.35%), dan terpinene 4 ol (10.63%). Jeruk purut memiliki efektivitas sebagai bakterisidal pada bakteri Propionibacterium acnes, 20 serotipe dari Salmonella dan juga bakteri lain yang dapat menyebabkan penyakit pada kulit seperti Staphylococcus epidermis dan Staphylococcus aureus. Jerawat atau acne vulgaris merupakan peradangan yang disertai penimbunan bahan keratin yang disebabkan karena adanya bakteri Staphylococcus aureus yang menyebabkan penyumbatan pada polisebasea. Tujuan penelitian ini adalah menformulasikan masker gel peel-off yang memiliki potensi anti bakteri Staphylococcus aureus sebagai salah satu pemicu timbulnya jerawat serta menguji sifat fisik sediaan. Formula gel peel-off dilakukan uji terhadap bakteri pada konsentrasi 10%, 15%, 20%, dan 25%. Evaluasi formula mencakup organoleptis, uji homogenitas, uji pH, uji viskositas, kemampuan untuk mengering, dan uji daya sebar. Hasil dari penelitian ini menunjukkan diameter zona hambat pada konsentrasi ekstrak kulit jeruk purt adalah sebesar 10% (1,0 cm), 15% (1,4 cm), 20% (1,7 cm), dan 25% (1,9 cm), kontrol positif (1,5 cm), kontrol negatif (0 cm). Meningkatnya konsentrasi menunjukkan meningkatnya kemampuan penghambatan terhadap Staphylococcus aureus serta uji fisik sediaan sesuai dengan literatur

The NEDD8 E3 ligase DCNL5 is phosphorylated by IKK alpha during Toll-like receptor activation

The activity of Cullin-RING ubiquitin E3 ligases (CRL) is regulated by NEDD8 modification. DCN-like proteins promote Cullin neddylation as scaffold-like E3s. One DCNL, DCNL5, is highly expressed in immune tissue. Here, we provide evidence that DCNL5 may be involved in innate immunity, as it is a direct substrate of the kinase IKKα during immune signalling. We find that upon activation of Toll-like receptors, DCNL5 gets rapidly and transiently phosphorylated on a specific N-terminal serine residue (S41). This phosphorylation event is specifically mediated by IKKα and not IKKβ. Our data for the first time provides evidence that DCNL proteins are post-translationally modified in an inducible manner. Our findings also provide the first example of a DCNL member as a kinase substrate in a signalling pathway, indicating that the activity of at least some DCNLs may be regulated

A single MIU motif of MINDY-1 recognizes K48-linked polyubiquitin chains

The eight different types of ubiquitin (Ub) chains that can be formed play important roles in diverse cellular processes. Linkage‐selective recognition of Ub chains by Ub‐binding domain (UBD)‐containing proteins is central to coupling different Ub signals to specific cellular responses. The motif interacting with ubiquitin (MIU) is a small UBD that has been characterized for its binding to monoUb. The recently discovered deubiquitinase MINDY‐1/FAM63A contains a tandem MIU repeat (tMIU) that is highly selective at binding to K48‐linked polyUb. We here identify that this linkage‐selective binding is mediated by a single MIU motif (MIU2) in MINDY‐1. The crystal structure of MIU2 in complex with K48‐linked polyubiquitin chains reveals that MIU2 on its own binds to all three Ub moieties in an open conformation that can only be accommodated by K48‐linked triUb. The weak Ub binder MIU1 increases overall affinity of the tMIU for polyUb chains without affecting its linkage selectivity. Our analyses reveal new concepts for linkage selectivity and polyUb recognition by UBDs

Mechanism of activation and regulation of Deubiquitinase activity in MINDY1 and MINDY2

Of the eight distinct polyubiquitin (polyUb) linkages that can be assembled, the roles of K48-linked polyUb (K48-polyUb) are the most established, with K48-polyUb modified proteins being targeted for degradation. MINDY1 and MINDY2 are members of the MINDY family of deubiquitinases (DUBs) that have exquisite specificity for cleaving K48-polyUb, yet we have a poor understanding of their catalytic mechanism. Here, we analyze the crystal structures of MINDY1 and MINDY2 alone and in complex with monoUb, di-, and penta-K48-polyUb, identifying 5 distinct Ub binding sites in the catalytic domain that explain how these DUBs sense both Ub chain length and linkage type to cleave K48-polyUb chains. The activity of MINDY1/2 is inhibited by the Cys-loop, and we find that substrate interaction relieves autoinhibition to activate these DUBs. We also find that MINDY1/2 use a non-canonical catalytic triad composed of Cys-His-Thr. Our findings highlight multiple layers of regulation modulating DUB activity in MINDY1 and MINDY2

MINDY-1 is a member of an evolutionarily conserved and structurally distinct new family of Deubiquitinating enzymes

Deubiquitinating enzymes (DUBs) remove ubiquitin (Ub) from Ub-conjugated substrates to regulate the functional outcome of ubiquitylation. Here we report the discovery of a new family of DUBs, which we have named MINDY (motif interacting with Ub-containing novel DUB family). Found in all eukaryotes, MINDY-family DUBs are highly selective at cleaving K48-linked polyUb, a signal that targets proteins for degradation. We identify the catalytic activity to be encoded within a previously unannotated domain, the crystal structure of which reveals a distinct protein fold with no homology to any of the known DUBs. The crystal structure of MINDY-1 (also known as FAM63A) in complex with propargylated Ub reveals conformational changes that realign the active site for catalysis. MINDY-1 prefers cleaving long polyUb chains and works by trimming chains from the distal end. Collectively, our results reveal a new family of DUBs that may have specialized roles in regulating proteostasis

Assembly and structure of Lys³³-linked polyubiquitin reveals distinct conformations

Ubiquitylation regulates a multitude of biological processes and this versatility stems from the ability of ubiquitin (Ub) to form topologically different polymers of eight different linkage types. Whereas some linkages have been studied in detail, other linkage types including Lys(33)-linked polyUb are poorly understood. In the present study, we identify an enzymatic system for the large-scale assembly of Lys(33) chains by combining the HECT (homologous to the E6–AP C-terminus) E3 ligase AREL1 (apoptosis-resistant E3 Ub protein ligase 1) with linkage selective deubiquitinases (DUBs). Moreover, this first characterization of the chain selectivity of AREL1 indicates its preference for assembling Lys(33)- and Lys(11)-linked Ub chains. Intriguingly, the crystal structure of Lys(33)-linked diUb reveals that it adopts a compact conformation very similar to that observed for Lys(11)-linked diUb. In contrast, crystallographic analysis of Lys(33)-linked triUb reveals a more extended conformation. These two distinct conformational states of Lys(33)-linked polyUb may be selectively recognized by Ub-binding domains (UBD) and enzymes of the Ub system. Importantly, our work provides a method to assemble Lys(33)-linked polyUb that will allow further characterization of this atypical chain type

K29-selective ubiquitin binding domain reveals structural basis of specificity and heterotypic nature of K29 polyubiquitin

Polyubiquitin chains regulate diverse cellular processes through the ability of ubiquitin to form chains of eight different linkage types. Although detected in yeast and mammals, little is known about K29-linked polyubiquitin. Here we report the generation of K29 chains in vitro using a ubiquitin chain-editing complex consisting of the HECT E3 ligase UBE3C and the deubiquitinase vOTU. We determined the crystal structure of K29-linked diubiquitin, which adopts an extended conformation with the hydrophobic patches on both ubiquitin moieties exposed and available for binding. Indeed, the crystal structure of the NZF1 domain of TRABID in complex with K29 chains reveals a binding mode that involves the hydrophobic patch on only one of the ubiquitin moieties and exploits the flexibility of K29 chains to achieve linkage selective binding. Further, we establish methods to study K29-linked polyubiquitin and find that K29 linkages exist in cells within mixed or branched chains containing other linkages

PEMBERDAYAAN IBU-IBU DASA WISMA SUKUN DALAM PEMBUATAN BATIK DENGAN TEKNIK ECOPRINT

Indonesia's large variety of natural ingredients makes Indonesia rich in a culture based on natural ingredients. Batik in Indonesia is an ancestral heritage that has existed from the time of the Hindu kingdom until now. The development of batik is increasing rapidly because market demand is always high. Natural fabric coloring with the ecoprint technique is an alternative to environmentally friendly raw materials and enhances the development of Indonesian batik. Ecoprint is a technique of natural dyeing in the form of flowers, leaves, stems, and bark on natural fibrous fabrics with the help of chemicals to bind natural colors into the fabric. The ecoprint technique is in the form of an iron blanket and a punch. This activity aims to provide knowledge and train the skills ecoprinting techniques of housewife communities in Dasa Wisma Sukun, Jetis, Wedomartani for batik production. The stages of this activity include the presentation of ecoprint, training and practice of making batik using ecoprint techniques, and monitoring and evaluating activities. The results from this activity are that partners have knowledge of ecoprint techniques and can implement ecoprint techniques in the production of batik cloth. In addition, partners are also more creative in using other natural ingredients such as natural dyes in the ecoprint technique. --- Beraneka ragamnya bahan alam di Indonesia menjadikan Indonesia kaya akan budaya berbasis bahan alam. Batik di Indonesia merupakan warisan leluhur yang sudah ada dari jaman kerajaan Hindu hingga sekarang. Perkembangan batik semakin pesat dikarenakan tingkat permintaan pasar yang selalu tinggi. Pewarnaan kain alami dengan teknik ecoprint menjadi salah satu alternatif pemanfaatan bahan alam yang ramah lingkungan dan meningkatkan perkembangan batik Indonesia. Ecoprint merupakan pengembangan pewarnaan kain dengan pewarna alami berupa bunga, daun, batang, kulit kayu pada kain berserat alami dengan bantuan zat kimia untuk mengikat warna alami ke dalam kain. Teknik ecoprint dibedakan menjadi 2 macam yaitu iron blanket dan pounding. Kegiatan ini bertujuan untuk memberikan pengetahuan dan melatih keterampilan ibu-ibu dasa wisma Sukun, Jetis, Wedomartani dalam teknik ecoprinting untuk produksi kain batik. Tahapan kegiatan ini antara lain pemaparan materi tentang ecoprint, pelatihan dan praktik pembuatan batik dengan teknik ecoprint, serta monitoring dan evaluasi kegiatan. Hasil yang dicapai dari kegiatan ini yaitu dari mitra memiliki pengetahuan tentang teknik ecoprint dan dapat mengimplementasikan teknik ecoprint dalam produksi kain batik. Selain itu, mitra juga menjadi lebih kreatif dalam menggunakan bahan alam lain sebagai pewarna alami dalam teknik ecoprint

DCNL5 is phosphorylated on Serine 41 by IKKα.

<p><b>(A</b>) Immunoblot analysis with the indicated antibodies of cell lysates from different cell lines. DCNL5 is strongly expressed in RAW264.7 (immortalized macrophages), Jurkat (immortalized T lymphocytes), and Raji (derived B lymphocytes) cells, while it is present at low levels in Thp1 cells (a monocyte-derived cell line). <b>(B</b>) Upper panel: Mass spectrum showing phosphopeptide precursor ions corresponding to DCNL5 (amino acids 39–48), which is phosphorylated at Ser41 in RAW264.7 macrophages treated with Poly (I:C) or Pam3CSK4, pre-treatment with MRT67307 (1 μM) for 1 hour enhance the phosphorylation. The results for unstimulated RAW264.7 macrophages are presented in blue (light), results for RAW264.7 macrophages stimulated with Poly (I:C) or Pam3CSK4 are in green (medium) and the results for RAW264.7 macrophages pre-treated with MRT67307 prior to stimulation with Poly (I:C) or Pam3CSK4 are depicted in red (heavy). For the Poly (I:C) treated samples, the ratio of labelled phosphopeptides in the different conditions quantified from MaxQUANT are given. Lower panel: Diagram of DCNL5 depicting the conserved C-ter PONY domain, its specific N-ter domain and the phosphorylated residue (red font; S41) uncovered by MS analysis. <b>(C)</b> Top panel: 1.4 μg of the purified recombinant DCNL5 was incubated with IKKα or IKKβ in the presence of <b>γ</b>32-ATP at 30°C for the times indicated. 1.4 μg of Iκbα was included as a positive control for IKKα and IKKβ. The stopped reactions were run on a gel and exposed to X-ray films. Right panel: same reactions as the top panel using IKKα WT or kinase dead mutant (S176A/S180A). The asterisk depicts degradation products from Iκbα that run at the same position as DCNL5. <b>(D)</b> <i>In vitro</i> phosphorylated DCNL5 was tryptic digested and separated by reverse chromatography followed by radioactivity measurement. Fractions corresponding to DCNL5 phosphopeptide were subjected for Edman sequencing. Flag tagged recombinant DCNL5 WT or S41A mutant was phosphorylated <i>in vitro</i> by IKKα WT in the presence of <b>γ</b>32-ATP at 30 °C for 30 minutes.</p