Immunoreactive glandular kallikrein in rat plasma: a radioimmunoassay for its determination
A radioimmunoassay (RIA) has been developed to measure immunoreactive glandular kallikrein in rat plasma. To prevent the binding of radioactive kallikrein to plasma inhibitors, /sup 125/I-kallikrein was inactivated with phenylmethylsulfonyl fluoride (PMSF), a procedure that maintained /sup 125/I-kallikrein immunoreactivity. Different volumes of plasma displaced /sup 125/I-PMSF-kallikrein in a parallel fashion to the kallikrein standard curve. The sensitivity of the RIA was 200 pg, and the recovery of nonradioactive active kallikrein added to plasma was 58.7%. The concentration of immunoreactive glandular kallikrein in normal rat plasma averaged 47.1 +/- 1.7 (SE) ng/ml. Bilateral nephrectomy caused a threefold increase in circulating glandular kallikrein (50 +/- 2.7 to 167 +/- 7 ng/ml; P < 0.001). Removal of the submandibular and sublingual glands signficantly decreased its concentration from 52 +/- 2.3 to 34 +/- 1.6 ng/ml (P < 0.001). Immunoreactive glandular kallikrein was higher in the submandibular gland vein than in arterial blood (venous; 94 +/- 10.5; arterial: 64 +/- 6.3 ng/ml; P < 0.05) and was lower in the renal venous blood (venous: 44 +/- 2.2; arterial: 53 +/- 2.6 ng/ml; P < 0.05). In conclusion, this study shows that the use of /sup 125/I-PMSF-kallikrein as tracer prevents the interference in the RIA caused by plasma protease inhibitors. It also indicates that the submandibular gland is an important source of the immunoreactive glandular kallikrein in rat plasma and that the kidney probably participates in its metabolism. Glandular kallikrein released by the submandibular gland into the circulation may participate in regulating local blood flow before it is inactivated by plasma inhibitors.
- Research Organization:
- Henry Ford Hospital, Detroit, MI
- OSTI ID:
- 5451100
- Journal Information:
- Am. J. Physiol.; (United States), Vol. 242:4
- Country of Publication:
- United States
- Language:
- English
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550601* - Medicine- Unsealed Radionuclides in Diagnostics