title: Characterization of the serpin, alpha 1-antichymotrypsin, in normal human cerebrospinal fluid.
abstract: Cerebrospinal fluid (CSF) from 20 male patients with nonneurologic disease (age 64.5 +/- 2.8 SEM) was analyzed for the presence of the serpin alpha 1-antichymotrypsin (alpha 1-ACT). A chymotrypsin-specific chromogenic substrate (succinyl-Ala-Ala-Pro-Phe-p-nitroanilide) was used to examine the CSF samples. All CSF samples showed inhibitory activity ranging from 45 to 80% inhibition. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the samples revealed the presence of a 68-kDa protein migrating identical to authentic human plasma alpha 1-ACT. Complex formation was performed with iodinated bovine chymotrypsin for several representative CSF samples having the highest chymotrypsin inhibitory activity. Comparison was made with complex formation performed with commercially available authentic human plasma alpha 1-ACT. These studies showed the formation of complexes at 37 degrees C, regardless of whether the sample was subsequently boiled or not. In the case of CSF, two complex bands, mass smaller than with plasma alpha 1-ACT, were formed at the lower temperature whereas a single higher Mr band was formed when the samples were boiled. To determine whether cleavage of the serpin occurred, these studies were repeated using human neutrophil cathepsin G as target protease. A complex of approximately 90 kDa was formed with human alpha 1-ACT under these same conditions. alpha 1-ACT has been detected in senile amyloid plaques in brains of Alzheimer's disease patients, the only plasma serine protease inhibitor localized to these structures. Another serpin, protease nexin I, is also found in these plaques, but this inhibitor does not circulate in plasma.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Neurobiology Research Laboratory, Department of Veterans Affairs Medical Center, Kansas City, Missouri 64128.
authors.forename: A
authors.initials: A
authors.lastname: Rayford
authors.forename: J S
authors.initials: JS
authors.lastname: Rao
authors.forename: B W
authors.initials: BW
authors.lastname: Festoff
mesh_terms: D000328:Adult; D015153:Blotting, Western; D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008297:Male; D011233:Precipitin Tests; D012016:Reference Values; D015843:Serpins / Q000134:cerebrospinal fluid*; D000514:alpha 1-Antichymotrypsin / Q000134:cerebrospinal fluid*
chemical_list: D015843:Serpins; D000514:alpha 1-Antichymotrypsin; D002918:Chymotrypsin