N-glycans of human protein C inhibitor: tissue-specific expression and function.
Protein C inhibitor (PCI) is a serpin type of serine protease inhibitor that is found in many tissues and fluids in human, including blood plasma, seminal plasma and urine. This inhibitor displays an unusually broad protease specificity compared with other serpins. Previous studies have shown that the N-glycan(s) and the NH₂-terminus affect some blood-related functions of PCI. In this study, we have for the first time determined the N-glycan profile of seminal plasma PCI, by mass spectrometry. The N-glycan structures differed markedly compared with those of both blood-derived and urinary PCI, providing evidence that the N-glycans of PCI are expressed in a tissue-specific manner. The most abundant structure (m/z 2592.9) had a composition of Fuc₃Hex₅HexNAc₄, consistent with a core fucosylated bi-antennary glycan with terminal Lewis(x). A major serine protease in semen, prostate specific antigen (PSA), was used to evaluate the effects of N-glycans and the NH₂-terminus on a PCI function related to the reproductive tract. Second-order rate constants for PSA inhibition by PCI were 4.3±0.2 and 4.1±0.5 M⁻¹ s⁻¹ for the natural full-length PCI and a form lacking six amino acids at the NH₂-terminus, respectively, whereas these constants were 4.8±0.1 and 29±7 M⁻¹ s⁻¹ for the corresponding PNGase F-treated forms. The 7-8-fold higher rate constants obtained when both the N-glycans and the NH₂-terminus had been removed suggest that these structures jointly affect the rate of PSA inhibition, presumably by together hindering conformational changes of PCI required to bind to the catalytic pocket of PSA.
Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden.
Wei
W
Sun
Paola
P
Grassi
Ake
A
Engström
Sanjeewani
S
Sooriyaarachchi
Wimal
W
Ubhayasekera
Julius
J
Hreinsson
Kjell
K
Wånggren
Gary F
GF
Clark
Anne
A
Dell
Sophia
S
Schedin-Weiss
D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D008958:Models, Molecular; D009928:Organ Specificity; D011134:Polysaccharides; D017430:Prostate-Specific Antigen / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D018046:Protein C Inhibitor / Q000097:blood / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D011487:Protein Conformation; D012661:Semen / Q000378:metabolism
D011134:Polysaccharides; D018046:Protein C Inhibitor; C534934:SERPINA5 protein, human; D017430:Prostate-Specific Antigen