Characterization of a monoclonal antibody B1 that recognizes phosphorylated Ser-158 in the activation peptide region of human coagulation factor IX.

Blood coagulation factor IX (FIX) undergoes various post-translational modifications such as gamma-carboxylation and glycosylation. Non-phosphorylated recombinant FIX has been reported to rapidly disappear from plasma, indicating that phosphorylation of FIX plays an important role in the physiological activity of this coagulation factor. In this study, we characterized the human FIX activation peptide (AP) using a monoclonal antibody that recognizes phosphorylated Ser-158 in the AP region. Murine monoclonal antibody B1 against human FIX recognized FIX with an apparent K(d) value of 5 nm in the presence of Ca(2+) (EC(50) = 0.58 mm). B1 bound to the isolated AP of FIX and retained the Ca(2+) dependence of binding to the isolated AP. The deglycosylation of AP did not affect the binding of B1 to AP, while B1 failed to bind to recombinant AP expressed in Escherichia coli. MALDI-TOF mass spectrometry showed that the m/z of plasma-derived deglycosylated AP is 82.54 Da greater than that of recombinant AP. The binding ability of B1 to AP was lost by the dephosphorylation of plasma-derived AP. B1 bound to synthetic peptide AP-(5-19), including phosphoserine-13, but not to the non-phosphorylated AP-(5-19) in the presence of Ca(2+). These data provide direct evidence that Ser-13 of the plasma-derived FIX AP region (Ser-158 of FIX) is phosphorylated and that B1 recognizes the epitope, which includes Ca(2+)-bound phosphoserine-158. B1 should be useful in the quality control of biologically active recombinant FIX containing phosphoserine-158.

Paleodemography of a medieval population in Japan: analysis of human skeletal remains from the Yuigahama-minami site.

The purpose of this study is to obtain demographic data regarding the medieval population buried at the Yuigahama-minami site in Kamakura, Japan, and to detect a secular trend in the life expectancy of Japanese population over the last several thousand years. The Yuigahama-minami skeletal sample consists of 260 individuals, including 98 subadults (under 20 years old) and 162 adults. A Yuigahama-minami abridged life-table analysis yielded a life expectancy at birth (e0) of 24.0 years for both sexes, a life expectancy at age 15 years (e15) of 15.8 years for males, and an e15 of 18.0 years for females. The reliability of the estimated e0 was confirmed by analysis of the juvenility index. Demographic profiles comparing the Yuigahama-minami series with other skeletal series indicated that both the survivorship curve and life expectancy of the Yuigahama-minami sample are similar to those of the Mesolithic-Neolithic Jomon population, but are far lower than those of the early modern Edo population. These comparisons strongly suggest that life expectancy changed little over the thousands of years between the Mesolithic-Neolithic Jomon and medieval periods, but then improved remarkably during the few hundred years between the medieval period and early modern Edo period. The short-lived tendency of the Yuigahama-minami sample does not contradict the archaeological hypothesis of unsanitary living conditions in medieval Kamakura. This is the first investigation to address the demographic features of a medieval population in Japan, and will help refine our understanding of long-term trends in the demographic profiles of inhabitants of Japan.