The tumor suppressor maspin does not undergo the stressed to relaxed transition or inhibit trypsin-like serine proteases. Evidence that maspin is not a protease inhibitory serpin.
The role of tumor suppressor proteins in the development of malignancy has made the understanding of their molecular mechanisms of action of great importance. Maspin is a tumor suppressor produced by a number of cell types of epithelial origin. Exogenous recombinant maspin has been shown to block the growth, motility, and invasiveness of breast tumor cell lines in vitro and in vivo. Although belonging to the the serine proteinase inhibitor (serpin) superfamily of proteins, the molecular mechanism of maspin is currently unknown. Here we show that the reactive site loop of maspin exists in an exposed conformation that does not require activation by cofactors. The reactive site loop of maspin, however, does not act as an inhibitor of proteinases such as chymotrypsin, elastase, plasmin, thrombin, and trypsin but rather as a substrate. Maspin is also unable to inhibit tissue and urokinase type plasminogen activators. Stability studies show that maspin cannot undergo the stressed-relaxed transition typical of proteinase-inhibitory serpins, and the protein is capable of spontaneous polymerization induced by changes in pH. It is likely, therefore, that maspin is structurally more closely related to ovalbumin and angiotensinogen, and its tumor suppressor activity is independent of a latent or intrinsic trypsin-like serine proteinase-inhibitory activity.
Pemberton
P A
PA
LXR Biotechnology, Richmond, California 94804, USA.
Wong
D T
DT
Gibson
H L
HL
Kiefer
M C
MC
Fitzpatrick
P A
PA
Sager
R
R
Barr
P J
PJ
D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000737:chemistry*; D005109:Extracellular Matrix / Q000378:metabolism; D016147:Genes, Tumor Suppressor; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D015843:Serpins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D000970:Antineoplastic Agents; D011506:Proteins; D011994:Recombinant Proteins; C085344:SERPIN-B5; D015843:Serpins