Mechanism of fibril formation by a 39-residue peptide (PAPf39) from human prostatic acidic phosphatase.
PAPf39 is a 39-residue peptide fragment from the sequence of human prostatic acidic phosphatase. This peptide was shown to form amyloid-like fibrils, which have been implicated in facilitating semen-mediated HIV transmission. Thus understanding molecular details of PAPf39 peptide fibril formation may aid in elucidating the mechanism of how PAPf39 fibrils are involved in HIV etiology. To this end, the kinetics of PAPf39 peptide fibrillization was studied using a battery of biophysical methods (atomic force microscopy, ThT fluorescence assays, far-UV circular dichroism spectroscopy, deep-UV resonance Raman spectroscopy, size exclusion chromatography, analytical ultracentrifugation, and small-angle X-ray scattering). It has been shown that fibril formation follows a nucleation-dependent elongation mechanism. Several critical factors for fibrillization have been identified. It was shown that agitation and/or seeding is required for fibril formation at 37 degrees C and neutral pH, with an additional requirement of a salt concentration above approximately 100 mM. Fibril formation by the PAPf39 peptide is inhibited by low pH or by low salt concentration at neutral pH. These observations suggest that the nucleation and fibrillization of the PAPf39 peptide are a tug-of-war between the interactions formed upon agitation and the electrostatic interactions, modulated by pH and salt concentration.
Ye
Zhuqiu
Z
Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York12065, USA.
French
Kinsley C
KC
Popova
Ludmila A
LA
Lednev
Igor K
IK
Lopez
Maria M
MM
Makhatadze
George I
GI
D000135:Acid Phosphatase; D000595:Amino Acid Sequence; D000682:Amyloid / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002850:Chromatography, Gel; D002942:Circular Dichroism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D018625:Microscopy, Atomic Force; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry / Q000378:metabolism*; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D013059:Spectrum Analysis, Raman; D013696:Temperature; D014461:Ultracentrifugation; D014965:X-Rays
D000682:Amyloid; D010446:Peptide Fragments; D000135:Acid Phosphatase; C428439:prostatic acid phosphatase; D017027:Protein Tyrosine Phosphatases