To assay for sample recovery of proteins during dialysis and fractionation, we analyzed sample aliquots that had been taken during this procedure for hK5 and hK6 by the aforementioned ELISA assays.
TOTAL PROTEIN, hK5, AND hK6 CONCENTRATIONS IN ROLLER BOTTLE CULTURE OVER TIME
This hormonal intervention produced a significant up-regulation of serum hK2, hK3, hK5, hK6
, hK7, and hK10 concentrations, with the most profound impact on hK2 and hK3.
Serum hK3 no longer showed a correlation with hK2 but still showed positive correlations with hK6
Because the hK4, hK6
, hK8, and hK11 proteins are produced by more restricted tissues (12, 21-23), hK7 might play more general roles than other human kallikreins, including the degradation of adhesion molecules between cells (24).
is recognized as a serine protease because the amino acid residues known to be crucial for substrate binding, specificity, and catalysis of the serine proteases (including serine, histidine, and aspartic acid) are highly conserved (5-8).
Recombinant hK1, hK2, hK3, hK4, hK5, hK6
, hK7, hK8, hK9, hK10, hK11, hK12, hK14, and hK15 proteins did not produce measurable readings, even at concentrations 1000-fold higher than that of hK13.
Because the KLK8 gene is predicted to encode for a secreted serine protease, this enzyme may have value as a biomarker, similar to hK3 (PSA) (3), hK6
, hK10 (NES1), and hK11 (TLSP) (4-7).
In addition to hK3 and hK2, hK6
and hK10 are emerging diagnostic markers for ovarian cancer (67-70).
The cross-reactivity of our assay with other kallikreins was determined by the use of purified, recombinant hK2 (250 [micro]g/L), hK3 (PSA; 1000 [micro]g/L), hK5 (1000 [micro]g/L), hK6
(2500 [micro]g/L), hK7 (1000 [micro]g/L), hK8 (1000 [micro]g/L), hK9 (1000 [micro]g/L), hK10 (1000 [micro]g/L), hK11 (1000 [micro]g/L), hK12 (1000 [micro]g/L), hK13 (1000 [micro]g/L), and hK14 (1000 [micro]g/L).
The cross-reactivities of other homologous proteins were investigated using purified recombinant PSA, hK2, and hK6
New gene Previous gene New protein symbol (a,b) symbol(s) symbol KLK1 KLK1 hK1 KLK3 KLK3 hK3 KLK2 KLK2 hK2 KLK4 PRSS17, KLK-L1, KLK4 hK4 KLK5 KLK-L2 hK5 KLK6 PRSS9 hK6
KLK7 PRSS6 hK7 KLKS PRSS19 hK8 KLK9 KLK-L3 hK9 KLK10 PRSSL1, hK10 KLK11 PRSS20 hK11 KLK12 KLK-L5 hK12 KLK13 KLK-L4 hK13 KLK14 KLK-L6 hK14 New gene symbol (a,b) Other protein names/symbols KLK1 Pancreatic/renal kallikrein, hPRK KLK3 Prostate-specific antigen, PSA KLK2 Human glandular kallikrein 1, hGK-1 KLK4 Prostase, KLK-L1 protein, EMSP1 KLK5 KLK-L2 protein, HSCTE KLK6 Zyme, protease M, neurosin KLK7 HSCCE KLKS Neuropsin, ovasin, TADG-14 KLK9 KLK-L3 protein KLK10 NES1 protein KLK11 TLSP/hippostasin KLK12 KLK-L5 protein KLK13 KLK-L4 protein KLK14 KLK-L6 protein New gene GenBank symbol (a,b) Accession No.