Cryptic origin of SPAI, a plasma protein with a transglutaminase substrate domain and the WAP motif, revealed by in situ hybridization and immunohistochemistry.
SPAI, originally isolated as a sodium/potassium-ATPase inhibitor and now considered to be a proteinase inhibitor of unknown specificity based on its similarity to elafin (an elastase inhibitor), is a new type of plasma protein that has a transglutaminase substrate domain, which serves as an anchoring sequence to be covalently cross-linked at target sites. To determine the source of SPAI, we carried out in situ hybridization and immunohistochemistry using an antisense cRNA probe and an antiserum against recombinant SPAI, respectively. Since previous RNase protection analysis had indicated that SPAI mRNA is almost exclusively expressed in the porcine small intestine, we used its frozen sections for the staining. The lower crypt was decorated with both the cRNA probe and antiserum, indicating that SPAI is synthesized and secreted by the enteroendocrine cells located near the crypt base. The native form of SPAI was also characterized by Western blotting. This result together with the previous biochemical and molecular biological characterizations may set the stage for identifying the physiological roles of the conceptually very interesting protein SPAI.
Furukawa
M
M
Department of Biological Sciences, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226, Japan.
Suzuki
Y
Y
Ghoneim
M A
MA
Tachibana
S
S
Hirose
S
S
D000818:Animals; D001798:Blood Proteins / Q000378:metabolism*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007413:Intestinal Mucosa / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors*; D013379:Substrate Specificity; D013552:Swine; D011503:Transglutaminases / Q000378:metabolism*
D001798:Blood Proteins; D011506:Proteins; D011503:Transglutaminases; D000254:Sodium-Potassium-Exchanging ATPase