Heterogeneity of hairy cell tartrate-resistant acid phosphatase.
The human nonerythrocytic acid phosphatases (AcP) are composed of seven distinct activity bands in nondenaturing polyacrylamide gel electrophoresis (PAGE) when stained using either 1-naphthyl phosphate or naphthol ASBI phosphate as substrate. They are numbered 0, 1, 2, 3, 3b, 4, and 5 according to their increasing mobility toward the cathode in acidic conditions. Of these, only the most cationic "band 5" is tartrate resistant (TRAcP). When naphthol ASBI phosphate is used as substrate, AcP activity can also be stained in situ. In the presence of tartrate, activity remains strong in the hairy cells (HC) of hairy cell leukemia (HCL). Thus, the TRAcP stain has remained a reliable marker for HC. To investigate the function of TRAcP in HC, we purified two isoforms of TRAcP from HCL spleen tissue and found them to have similar substrate specificities and inhibitor sensitivities. In this report, we describe in detail the methods for TRAcP purification and compare some of the structural properties of the two isoforms to reinforce the concept that human TRAcP is a heterogeneous group of related enzymes. Band 5 represented only 15-20% of the total TRAcP extracted from HCL spleen. The remaining 80% of TRAcP hydrolyzed p-nitrophenyl phosphate but not naphthol ASBI phosphate and was not detectable in acidic, nondenaturing PAGE gels. Band 5 was solubilized from tissue using 500 mmol/L NaCl after previous extraction with 0.5% (v/v) NP-40 removed most other AcP and TRAcP activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Janckila
A J
AJ
Medical Service, Department of Veterans Affairs Medical Center, Louisville, KY 40206.
Latham
M D
MD
Lam
K W
KW
Chow
K C
KC
Li
C Y
CY
Yam
L T
LT
D000135:Acid Phosphatase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D014408:Biomarkers, Tumor; D003124:Colorimetry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D007943:Leukemia, Hairy Cell / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013045:Species Specificity; D013154:Spleen / Q000201:enzymology*; D000071681:Tartrate-Resistant Acid Phosphatase; D013644:Tartrates / Q000494:pharmacology
D014408:Biomarkers, Tumor; D007527:Isoenzymes; D013644:Tartrates; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; C029768:tartaric acid