Comparison of the molecular forms of the Kex2/subtilisin-like serine proteases SPC2, SPC3, and furin in neuroendocrine secretory vesicles reveals differences in carboxyl-terminus truncation and membrane association.
The molecular forms and membrane association of SPC2, SPC3, and furin were investigated in neuroendocrine secretory vesicles from the anterior, intermediate, and neural lobes of bovine pituitary and bovine adrenal medulla. The major immunoreactive form of SPC2 was the full-length enzyme with a molecular mass of 64 kDa. The major immunoreactive form of SPC3 was truncated at the carboxyl terminus and had a molecular mass of 64 kDa. Full-length 86-kDa SPC3 with an intact carboxyl terminus was found only in bovine chromaffin granules. Immunoreactive furin was also detected in secretory vesicles. The molecular masses of 80 and 76 kDa were consistent with carboxyl-terminal truncation of furin to remove the transmembrane domain. All three enzymes were distributed between the soluble and membrane fractions of secretory vesicles although the degree of membrane association was tissue specific and, in the case of SPC3, dependent on the molecular form of the enzyme. Significant amounts of membrane-associated and soluble forms of SPC2, SPC3, and furin were found in pituitary secretory vesicles, whereas the majority of the immunoreactivity in chromaffin granules was membrane associated. More detailed analyses of chromaffin granule membranes revealed that 86-kDa SPC3 was more tightly associated with the membrane fraction than the carboxyl terminus-truncated 64-kDa form.
Hill
R M
RM
School of Biological Sciences, University of Auckland, New Zealand.
Ledgerwood
E C
EC
Brennan
S O
SO
Pu
L P
LP
Loh
Y P
YP
Christie
D L
DL
Birch
N P
NP
D000313:Adrenal Medulla / Q000378:metabolism; D000818:Animals; D043423:Carboxypeptidase H; D002268:Carboxypeptidases / Q000378:metabolism; D002417:Cattle; D002837:Chromaffin Granules / Q000378:metabolism; D045683:Furin; D007158:Immunologic Techniques; D008566:Membranes / Q000378:metabolism; D008968:Molecular Conformation; D009490:Neurosecretory Systems / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000378:metabolism*; D012995:Solubility; D013381:Subtilisins / Q000737:chemistry* / Q000378:metabolism*
D002268:Carboxypeptidases; D043423:Carboxypeptidase H; D012697:Serine Endopeptidases; D013381:Subtilisins; D045683:Furin