Properties of cannabinoid receptors in medicinal plant Cannabis sativa

Biljka Cannabis sativa, konoplja, sadrži mnoga ljekovita svojstva koja se polako otkrivaju. Nakon otkrića endokanabinoidnogasustava, ustanovljena je poveznica između ljudskoga tijela i biljke kanabis. Među mnogim medicinskim svojstvima, endokanabinoidni sustav pokazuje utjecaj na rast tumorskih stanica, njihovo širenje, angiogenezu, migraciju i temelj je budućih ciljnih istraživanja u farmakoterapiji. Također, brojne studije ukazuju na nove uvide u terapeutsko liječenje neurodegenerativnih bolesti poput multiple skleroze i Parkinsonove bolesti, glaukoma, gojaznosti i povezanih metaboličkih bolesti te raka. Anti-tumorske tvari ili kanabinoidi jesu supresori rasta tumora, te je poznato kako stimuliraju apetit u pacijenata oboljelih od raka ili HIV-a, te sprečavaju povraćanja i mučnine u kemoterapijskih pacijenata. Unatoč svemu, ta je biljka i dalje klasificirana kao narkotik. No, ono što čini razliku između zlouporabe konoplje i njezine medicinske upotrebe jesu konstruktivna znanstvena istraživanja i uravnotežena medicinska primjena. Ovaj članak pokriva mnoga svojstva i aspekte primjene konoplje, kako bi se omogućilo bolje razumijevanje te kontroverzne droge kao medicinske biljke.Cannabis plant (Cannabis sativa) contains many healing properties which are being slowly revealed. After the endocannabinoid system discovery, it has been acknowledged that human body is interacting with the cannabis. Among many medical aspects, the endocannabinoid system exerts effect on the growth of tumour cells, its proliferation, angiogenesis and migration and represents an emerging target of pharmacotherapy. Also, many studies have delivered new insights into the therapeutic targeting of this system for treating neurodegenerative disorders such as multiple sclerosis and Parkinson’s disease, glaucoma, obesity and related metabolic disorders, also cancer, etc. Effective antitumoural agents or cannabinoids are suppressors of tumour growth, also known for stimulating appetite in cancer and HIV affected patients, as well as for preventing vomiting and nausea in chemotherapy patients. Nevertheless, this plant is still classified as a narcotic; however what makes a distinction between drug abuse and medicine useare constructive and well-designed research investigations and balanced medical application. This article, hopefully, covers as many aspects of cannabis as possible, in order to provide better understanding of this controversial drug as a medicine plant

Biological chips unlock the secrets of genome

Nedavna otkrića na području tehnologije DNA-čipova omogućuju znanstvenicima istraživanje ekspresije tisuća gena u jednoj jedinoj reakciji, pri čemu se stvara velika količina podataka. Biološki se čipovi temelje na metodi hibridizacije, te dozvoljavaju uvid u ekspresiju gena na razini čitavoga genoma. Štoviše, čipovi se upotrebljavaju kako bismo istražili stanične i signalne putove i osnovne aspekte rasta i razvoja organizma tj. biološke životne procese. Nedavno sekvencionirani genomi biljke Arabidopsis thaliana i čovjeka otkrivaju postojanje mnoštva gena uključenih u određene ljudske bolesti koji imaju svoje ortologe prisutne u višim biljkama. Ključni proteini uključeni u bolesti, a posebno njihovi načini ekspresije i principi nastalih mutacija, sve se više objašnjavaju primjenom čipova. Navedeni će tehnološki napredak, nadamo se, pridonijeti skorom otkriću podrijetla genetskih bolesti i ubrzati stvaranje specifičnih genskih terapija, a čemu ćemo barem malim djelićem pridonijeti i mi našim istraživanjima.Recent advances in microarray technology allowed scientists to investigate expression of thousands of genes in a single reaction, thus generating a vast amount of data. Biological chips, based on hybridization methods, enable detection of genes on a genomic scale. Moreover, chips are used to elucidate fundamental life processes such as cellular and signalling pathways and basic aspects of growth and development of organisms. Recently sequenced model plant Arabidopsis thaliana and human genome revealed that a majority of human genes involved directly or indirectly in certain human diseases have their orthologs in vacular plants. Determination of key proteins involved in human health and disease, especially their gene expression patterns and mutations, are becoming rapidly examined and investigated by using biological chips. These technological advances will hopefully uncover the origin of genetic illnesses and likely lead to creation of specific gene therapies

The first dipeptidyl peptidase III from a thermophile: Structural basis for thermal stability and reduced activity

Dipeptidyl peptidase III (DPP III) isolated from the thermophilic bacteria Caldithrix abyssi (Ca) is a two-domain zinc exopeptidase, a member of the M49 family. Like other DPPs III, it cleaves dipeptides from the N-terminus of its substrates but differently from human, yeast and Bacteroides thetaiotaomicron (mesophile) orthologs, it has the pentapeptide zinc binding motif (HEISH) in the active site instead of the hexapeptide (HEXXGH). The aim of our study was to investigate structure, dynamics and activity of CaDPP III, as well as to find possible differences with already characterized DPPs III from mesophiles, especially B. thetaiotaomicron. The enzyme structure was determined by X-ray diffraction, while stability and flexibility were investigated using MD simulations. Using molecular modeling approach we determined the way of ligands binding into the enzyme active site and identified the possible reasons for the decreased substrate specificity compared to other DPPs III. The obtained results gave us possible explanation for higher stability, as well as higher temperature optimum of CaDPP III. The structural features explaining its altered substrate specificity are also given. The possible structural and catalytic significance of the HEISH motive, unique to CaDPP III, was studied computationally, comparing the results of long MD simulations of the wild type enzyme with those obtained for the HEISGH mutant. This study presents the first structural and biochemical characterization of DPP III from a thermophile

Interdisciplinary study of the effects of dipeptidyl-peptidase III cancer mutations on the KEAP1-NRF2 signaling pathway

Dipeptidyl peptidase III (DPP III) is associated with cancer progression via interaction with KEAP1, leading to upregulation of the KEAP1-NRF2 oxidative stress pathway. Numerous DPP III mutations have been found in human tumor genomes, and it is suggested that some of them may alter affinity for KEAP1. One such example is the DPP III-R623W variant, which in our previous study showed much higher affinity for the Kelch domain of KEAP1 than the wild-type protein. In this work, we have investigated the effects of this mutation in cultured cells and the effects of several other DPP III mutations on the stability of KEAP1-DPP III complex using an interdisciplinary approach combining biochemical, biophysical and molecular biology methods with computational studies. We determined the affinity of the DPP III variants for the Kelch domain experimentally and by molecular modeling, as well as the effects of the R623W on the expression of several NRF2-controlled genes. We confirmed that the R623W variant upregulates NQO1 expression at the transcriptional level. This supports the hypothesis from our previous study that the increased affinity of the R623W variant for KEAP1 leads to upregulation of the KEAP1-NRF2 pathway. These results provide a new perspective on the involvement of DPP III in cancer progression and prognosis