TY - JOUR
T1 - Inhibition of the cysteine proteinases cathepsins K and L by the serpin headpin (SERPINB13)
T2 - A kinetic analysis
AU - Jayakumar, Arumugam
AU - Kang, Ya'an
AU - Frederick, Mitchell J.
AU - Pak, Stephen C.
AU - Henderson, Ying
AU - Holton, Paula R.
AU - Mitsudo, Kenji
AU - Silverman, Gary A.
AU - EL-Naggar, Adel K.
AU - Brömme, Dieter
AU - Clayman, Gary L.
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2003/1/15
Y1 - 2003/1/15
N2 - Headpin (SERPINB13) is a novel member of the serine proteinase inhibitor (Serpin) gene family that was originally cloned from a keratinocyte cDNA library. Western blot analysis using a headpin-specific antiserum recognized a protein with the predicted Mr of 44kDa in lysates derived from a transformed keratinocyte cell line known to express headpin mRNA. Similarity of the reactive-site loop (RSL) domain of headpin, notably at the P1-P1′ residues, with other serpins that inhibit cysteine and serine proteinases suggests that headpin may inhibit similar proteinases. This study demonstrates that recombinant headpin indeed inhibits cathepsins K and L, but not chymotrypsin, elastase, trypsin, subtilisin A, urokinase-type plasminogen activator, plasmin, or thrombin. The second-order rate constants (ka) for the inhibitory reactions of rHeadpin with cathepsins K and L were 5.1±0.6×104 and 4.1±0.8×104M-1s-1, respectively. Headpin formed SDS-stable complexes with cathepsins K and L, a characteristic property of inhibitory serpins. Interactions of the RSL domain of headpin with cathepsins K and L were indicated by cleavage of headpin near the predicted P1-P1′ residues by these proteinases. These results demonstrate that the serpin headpin possesses specificity for inhibiting lysosomal cysteine proteinases.
AB - Headpin (SERPINB13) is a novel member of the serine proteinase inhibitor (Serpin) gene family that was originally cloned from a keratinocyte cDNA library. Western blot analysis using a headpin-specific antiserum recognized a protein with the predicted Mr of 44kDa in lysates derived from a transformed keratinocyte cell line known to express headpin mRNA. Similarity of the reactive-site loop (RSL) domain of headpin, notably at the P1-P1′ residues, with other serpins that inhibit cysteine and serine proteinases suggests that headpin may inhibit similar proteinases. This study demonstrates that recombinant headpin indeed inhibits cathepsins K and L, but not chymotrypsin, elastase, trypsin, subtilisin A, urokinase-type plasminogen activator, plasmin, or thrombin. The second-order rate constants (ka) for the inhibitory reactions of rHeadpin with cathepsins K and L were 5.1±0.6×104 and 4.1±0.8×104M-1s-1, respectively. Headpin formed SDS-stable complexes with cathepsins K and L, a characteristic property of inhibitory serpins. Interactions of the RSL domain of headpin with cathepsins K and L were indicated by cleavage of headpin near the predicted P1-P1′ residues by these proteinases. These results demonstrate that the serpin headpin possesses specificity for inhibiting lysosomal cysteine proteinases.
KW - Cysteine proteinases
KW - Headpin
KW - Insect cells
KW - Proteinase inhibition
KW - Serine proteinase inhibitor
UR - http://www.scopus.com/inward/record.url?scp=0037440423&partnerID=8YFLogxK
U2 - 10.1016/S0003-9861(02)00635-5
DO - 10.1016/S0003-9861(02)00635-5
M3 - Article
C2 - 12504904
AN - SCOPUS:0037440423
VL - 409
SP - 367
EP - 374
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
SN - 0003-9861
IS - 2
ER -