• 목록
• 게시판 리스트 옵션
  • 수정
  • 삭제

Predict specificity for trypsin, but not chymotrypsin. In spite of this, chymotrypsin is inhibited by BPTI. The WFDC genes at the locus on chromosome 20 could be important during sperm maturation, as several of them are highly expressed in the epididymis. There is this far no report on target proteases in the epididymis. It has also been speculated that the WFDC genes are important but not fully recognized components, of innate immunity; in which some modulate host response by regulating endogenous pro-inflammatory proteases, such as elastase and cathepsin G, whereas others regulate poorly characterized exogenous proteases used for tissue invasion by microbes [16]. The genes expressing Kunitz domains could have a similar function. In this report it is shown that SPINT3 and SPINT4 with high specificity are expressed in the epididymis, which suggests a role in the maturation of spermatozoa. An innate immune function can also inferred, as it has been shown that the Kunitz domain of SPINLW1 inhibits elastase and display antibacterial activity [32].which the genes could have been formed, include an exon shuffling event, in which the Kunitz motif was taken up by a WFDC gene, and a following purifying process, in which the WFDC domain was lost. This would then be the third mechanism, beside de novo selection of splice site and repeat expansion, by which the regulatory frame of a conserved WFDC/semen coagulum gene could generate a new protein product. The restricted expression of the novel genes to the epididymis suggests that the secreted Kunitz domains are important in male reproduction. The recombinantly produced proteins are presumably bioactive, as demonstrated with similarly made BPTI, but may have a narrower spectrum of inhibition, as indicated by the lacking activity against eight PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/13485127 proteases with differing specificity. Another possibility is that they have lost the protease inhibiting property, which is typical of Kunitz domains, in favor of new hitherto unidentified functions.MethodsIdentification of novel genes in genomic DNA sequencesThe nucleotide sequence of the WFDC-locus on chromosome 20 was translated in six reading frames and scanned for short sequence motifs as previously described [14]. Kunitz domains were localized using the motif GCXGN and NNF (single letter amino Fmoc-Oic-OH acid code where ?means any residue) and confirmed by identification of appropriately spaced Cys residues through manual inspection of the sequence.RT-PCRConclusions We have in this report described three genes that produce small secreted proteins, composed of single Kunitz domains that are relatively specific for the epididymis. The similar organization, chromosomal location and site of expression, suggests that the novel genes encoding Kunitz motifs are homologous with the genes of WFDC-type protease inhibitors and semen coagulum proteins, in spite of lacking similarity in primary structure of their protein products. A likely mechanism, byPreparation of RNA samples and conditions for cDNA synthesis has been described earlier [15]. Primers were designed from nucleotide sequences originating from different exons to enable differentiation between PCR products from spliced transcripts and non-spliced PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/14445666 transcripts or genomic DNA (Table 4). A panel of cDNA derived from different tissues were screened by PCR using previously described conditions [14]. The PCR products were analyzed by 4-Bromopicolinaldehyde electrophoresis in 2 agarose gels stained by ethidium bromide. Samples that by thisClau.

• 페이스북 공유
• 트위터 공유
• 구글+ 공유