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Molecular Characterisation and Expression Analysis of
Cathepsin D
from the Asian Seabass Lates calcarifer
(Pencirian Molekul dan Analisis Pengekspresan Katepsin D daripada
Ikan Siakap Lates calcarifer)
SHARIZA AZIZAN1, KIEW-LIAN WAN1,2* & ADURA MOHD-ADNAN2
1School of Biosciences and Biotechnology, Faculty of Science and
Technology
Universiti Kebangsaan Malaysia, 43600 UKM Bangi,
Selangor D.E. Malaysia
2Malaysia Genome Institute, Jalan Bangi, 43000 Kajang, Selangor,
D.E. Malaysia
Diserahkan: 18 Mei 2013/Diterima: 26 November
2013
ABSTRACT
The lysosomal aspartic proteinase cathepsin D is an acute phase
protein involved in various physiological processes, including vitellogenesis,
yolk processing and immune responses. In this study, we characterised the
cathepsin D from the Asian seabass Lates calcarifer and examined its expression profile during
infection. The complete coding sequence of L. calcarifer cathepsin D
consists of 1191 nucleotides, encoding a 396 amino acid protein molecule that
is made up of a putative signal peptide, a leader peptide and a mature peptide.
Phylogenetic analyses showed that two types of cathepsin D are present in the
teleost lineage i.e. cathepsin D1 and D2, whereas higher vertebrates possess
only one type of cathepsin D. L. calcarifer cathepsin D was clustered
together with cathepsin D1 from other teleosts. Compared to mammalian
sequences, L. calcarifer cathepsin D lacks the β-hairpin loop that
forms the double chain and is present as a single chain peptide with conserved
aspartic active sites like other fish. Both multiple sequence alignment and
phylogenetic analysis indicated that the L. calcarifer cathepsin D
sequence codes for cathepsin D1 and suggested that it shares the same functions
with cathepsin D from other fish. Expression profiling analysis of cathepsin D
in L. calcarifer infected with Aeromonas hydrophila showed that
it is up-regulated in immune-related tissues such as gills, spleen and liver,
suggesting that cathepsin D plays an important role in the innate immune
response of L. calcarifer against pathogens.
Keywords: Acute phase protein; aspartic proteinase; expression
profile; innate immune response
ABSTRAK
Katepsin D, sejenis proteinase aspartik lisosom merupakan protein
fasa akut yang terlibat dalam pelbagai proses fisiologi, termasuk
vitelogenesis, pemprosesan yolka dan gerak balas keimunan. Dalam
kajian ini, kami telah mencirikan katepsin D daripada ikan siakap
Lates calcarifer dan mengkaji profil pengekspresannya
semasa infeksi. Jujukan pengekodan lengkap katepsin D L. calcarifer
terdiri daripada 1191 nukleotida yang mengekod molekul protein
bersaiz 396 asid amino yang merangkumi satu peptida isyarat putatif,
satu peptida mendahului dan satu peptida matang. Analisis filogenetik
menunjukkan bahawa terdapat dua jenis katepsin D hadir dalam susur
galur teleost iaitu katepsin D1 dan D2, sementara vertebrata peringkat
tinggi hanya mempunyai satu jenis katepsin D. Katepsin D L.
calcarifer dikelompokkan bersama dengan katepsin D1 ikan teleost
lain. Berbanding dengan jujukan mamalia, katepsin D L. calcarifer
didapati tidak mempunyai jujukan gelung pin rambut β
yang membentuk rantai ganda dua dan wujud sebagai rantai tunggal
peptida dengan kehadiran tapak aktif aspartik terpulihara seperti
ikan lain. Penjajaran jujukan berbilang dan analisis filogenetik
menunjukkan bahawa jujukan katepsin D L. calcarifer mengekodkan
katepsin D1 dan mencadangkan ia mempunyai
fungsi yang sama dengan katepsin D ikan lain. Analisis profil pengekspresan
katepsin D dalam L. calcarifer terinfeksi Aeromonas hydrophila
mendedahkan bahawa pengekspresannya meningkat dalam tisu berkait-keimunan
seperti insang, limpa dan hepar mencadangkan bahawa katepsin D memainkan
peranan yang penting dalam gerak balas keimunan semula jadi L.
calcarifer terhadap patogen.
Kata kunci: Gerak balas keimunan semula jadi;
profil pengeskpresan; proteinase aspartik; protein fasa akut
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*Pengarang untuk surat-menyurat; email: klwan@ukm.edu.my
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