Materials and Methods
We have done various deep analysis on human BLK sequence, homology
modelling is the development of an atomic version of a target protein
primarily based totally at the goal’s amino acid series and the
templates; experimentally determined systems of homologous proteins. In
recent years structural biology shifted towards structure-based drug
discovery. We have modelled the structure using the techniques of
homology modelling (Jacobson et al.,
2004; Waterhouse et al., 2018). The
structure was pre-processed using the protein preparation wizard and
have filled the gap using Prime, the further structure was minimized in
terms of its energy using the same wizard for the next step of the
analysis. The active site is the region of an enzyme in which substrate
molecules bind and go through a chemical reaction. The lively site
consists of residues that shape brief bonds with the binding site and
residues that catalyse a response of the catalytic site, prediction of
the active site of the BLKhave bee done using SiteMap
(Singh et al., 2011;
Wass et al., 2010) with the specific
parameters of more restrictive and standard grid of crops site of 4 maps
for the further analysis and have got total of 5 active site. We have
also extracted the amino acid distribution throughout the sequence using
the ProtParam and got the CSV format data after that we have performed
various analysis in terms of statistical analysis and have plotted the
data for the quick understanding of the BLK. ProtParam lets in the
computation of diverse bodily and chemical parameters such as molecular
weight, theoretical pI, amino acid composition, atomic composition,
extinction coefficient, estimated half-life, instability index,
aliphatic index and grand average of hydropathicity for a given protein
saved in Swiss-Prot or TrEMBL or for a user-entered protein
sequence(Gasteiger et al., 2005).
Secondary structure of the protein has been predicted using emboss
(Garnier et al., 1978). GeneCards server
was used for the categorisation of mRNA expression level in different
human tissues. Antigenic regions on protein defined a lot about
inhibitors and the future research scope on that particular protein. In
our studies Emboss was used for the extraction of the antigenic region
and Geneious prime was used for the plotting of the data. Additionally,
we have also used InterProScan for the characterisation of the domains
on the surface of BLK for better functional understanding. We have
collected BLK from thousands of species and have mined the best, based
on the characterisation and have taken the only largest size of BLK from
every species and got a total of 324 species. Further data have been
aligned using MAFFT v7.450 Alignment
(Rozewicki et al., 2019). We have used
FFT-NS-1 and 200PAM/k=2 as main principle parameters and have got
comparative annotations of every aspect of the various BLK. Geneious
Tree has performed for the plotting of the tree with automatic direction
determiner. Further, Itol server (Letunic
& Bork, 2019) used for editing of a phylogenetic tree for better
understanding with its annotation characterization and bootstrapping.