Gel Permeation and Ion-Exchange Chromatography of Proteins and Peptides

Kato, Simone Parvez, Hasan Parvez

• Publisher: CRC Press
• Year: 2020
• Print ISBN: 9789067640480
• eBook ISBN: 9781000083170
• Topic: Physical Sciences
• Subtopic: Occupational & Industrial Medicine

This book is a collection of critical reviews of the use of high-performance liquid chromatography in a very specialized area of research. It describes in detail modern methodology to separate nucleic acids, enzymes and a wide variety of biologically active proteins such as renin.

High-performance liquid chromatography for enkephalins and enkephalin-containing peptides

KOHICHI KOJIMA and TOSHIHARU NAGATSU

Laboratory of Cell Physiology, Department of Life Chemistry, Graduate School at Nagatsuta, Tokyo Institute of Technology, Yokohama 227, Japan

Introduction

The endogenous opiate peptides were first discovered and identified as penta-peptides, methionine-enkephalin (Tyr–Gly–Gly–Phe–Met, Met-enkephalin) and leucine-enkephalin (Tyr–Gly–Gly–Phe–Leu, Leu-enkephalin) in 1975 (Hughes et al., 1975), Met-enkephalin was subsequently found to be a part of a previously discovered pituitary peptide, β-lipotropin (β-LPH) with 91 amino acid residues (Li et al., 1965). The sequence from residues 61 to 65 of β-LPH is identical to Met-enkephalin. But no peptide containing the sequence for Leu-enkephalin was known at that time. Within a few years of the discovery of Met- and Leu-enkephalins in pig brain, three enkephalin-containing (EC-) proteins for Met-enkephalin and Leu-enkephalin have been identified as the possible precursors for opioid peptides [pre-proopiomelanocortin (Nakanishi et al., 1979), pre-proenkephalin (pre-pro-enkephalin A) (Noda et al., 1982; Gubler et al., 1982), and pre-prodynorphin (pre-proenkephalin B) (Kakidani et al., 1982)].

In this chapter, we review the identification and purification of EC-proteins and peptides, structure analysis, and their enzymatic processing by using high-performance liquid chromatography (HPLC).

Identification and Purification of Ec-Peptides and Proteins

Soon after the discovery of the two pentapeptides (Met-enkephalin and Leu-enkephalin), precursor EC-proteins were subsequently isolated from the pituitary gland of rat (Rubinstein et al., 1978) and camel (Kimura et al., 1979) using HPLC. Table 1 summarizes isolation and/or purification of EC-peptides and proteins using HPLC.

Table 1. Isolation and purification of enkephaltlin-containing peptides and proteins

Starting material	Found or isolated substance(s)	Column(s)	References
Porcine hypothalamus neurohypophysis	α-Endorphin and γ-Endorphin	μ-Bondapak C18	Ling et al. (1976)
Rat anterior pituitary	β-Lipotropin	Partisil SCX and LiChrosorb RP-18	Rubinstein et al. (1977a)
Rat pituitary	β-Endorphin	Partisil SCX and LiChrosorb RP-18	Rubinstein et al. (1977b)
Rat anterior pituitary	Pro-opiocortin	LiChrosorb PR-18	Rubinstein et al. (1978)
Camel pituitary	Pro-opiocortin	LiChrosorb RP-8	Kimura et al. (1979)
Bovine adrenal medulla and adrenal medullary chromaffin granules	Putative enkephalin precursors (mol. wt 20000, 10 000, and 5000)	LiChrosorb RP-18	Lewis et al. (1979)
Porcine pituitary	Dynorphin-(1-13)	Cig column	Goldstein et al. (1979)
Porcine hypothalami	Pro-methionine-enkephalin (H-Tyr-Gly-Gly-Phe-Met(O)-Arg-OH)	μBondapak C18	Huang et al. (1979)
Bovine adrenal chromaffin granules	Met-enkephalin[Arg^6, Phe^7] (Tyr-Gly-Gly-Phe-Met-Arg-Phe) and a few other peptides	LiChrosorb RP-18	Stern et al. (1979)
Bovine adrenal chromaffin granules	Peptides of 3000-5000 (one has two copies of Met-enkephalin sequence the other has one Met-enkephalin and one Leu-enkephalin)	LiChrosorb RP-18, Ultrasphere Octyl and Spherisorb CN	Kimura et al. (1980)
Bovine adrenal chromaffin granules	Proteins of 8000 and 14 000	LiChrosorb RP-18, Ultrasphere ODS, Spherisorb CN and diphenyl-RP	Lewis et al. (1980c)
Human adrenal medullary tumour	Met-enkephalin precursors (mol. wt ~ 2000)	Spherisorb ODS and Partisil ODS	Clement-Jones et al. (1980)
Bovine adrenal chromaffin granules, bovine striata and human putamen	Hexa- and Hepta- peptides (Met-enkephalin-Arg^6-Arg^7, Met-enkephalin-Arg^6 -Phe ^7, Met-enkephalin-Lys^6, Met-enkephalin-Arg^6 and Leu-enkephalin-Arg^6)	LiChrosorb RP-18, Ultrasphere ODS and diphenyl-RP	Stern et al. (1980)
Bovine adrenal medulla	Dodecapeptide (BAM-12P)	μBondapak C-18	Mizuno et al. (1980a)
Bovine adrenal medulla	Docosa- and Eicosa-peptide (BAM-22P and BAM-20P)	μBondapak C-18	Mizuno et al. (1980b)
Bovine adrenal chromaffin granules	3600 and 4900 enkephalin-containing polypeptides	LiChrosorb RP-18, Ultrasphere ODS and diphenyl-RP	Stern et al. (1981)
Bovine adrenal chromaffin granules	3 200 enkephalin-containing peptide (contains both a Met- and Leu-enkephalin sequence)	LiChrosorb RP-18, Ultrasphere ODS and diphenyl-RP	Kilpatrick et al. (1981a)
Porcine hypothalami	β-Neo-endorphin	μBondapak C-18	Minamino et al. (1981)
Porcine pituitary	Dynorphin-(1-17)	Reverse-phase column	Goldstein et al. (1981)
Bovine adrenal medulla chromaffin granules	Met-enkephalin-Arg^6 -Gly ^7 -Leu^8	LiChrosorb RP-18 and Ultrasphere ODS	Kilpatrick et al. (1981b)
Bovine adrenal medulla chromaffin granules	A 5300 enkephalin-containing polypeptide (contains single Met-enkephalin-Arg^6 -Gly ^7 -Leu^8 sequence)	LiChrosorb RP-18 and Zorbax TMS	Jones et al. (1982a)
Porcine duodenum	Dynorphin-(l-17)	Nucleosil C18 and Nucleosil Phenyl	Tachibana et al. (1982)
Bovine adrenal medulla chromaffin granules	18 200 enkephalin-containing polypeptide (contains four Met-enkephalin sequences)	LiChrosorb RP-8, LiChrosorb CN and diphenyl-RP	Kilpatrick et al. (1982a)
Bovine posterior pituitary glands	Rimorphin (contains one Leu-enkephalin sequence)	Ultrasphere ODS and Nucleosil phenyl	Kilpatrick et al. (1982b)
Porcine pituitary	A 4000 dynorphin	μBondapak C18	Fischli et al. (1982)
Bovine posterior pituitary glands	Rimorphin, dynorphin and α-neo-endorphin	Ultrasphere ODS Nucleosil phenyl	Kilpatrick et al. (1982c)
Bufo marinus brain	Enkephalins and proenkephalin-derived enkephalin-containing peptides	Nucleosil C18 and Nucleosil phenyl	Kilpatrick et al. (1983a)
Bovine caudate nucleus	Tyr-Gly-Gly-Phe-Met-Arg-Arg-Val-NH2	Ultrasphere ODS	Weber et al. (1983)

Before these experiments, smaller precursors, α and γ-endorphin (Ling et al., 1976), β-lipotropin (Rubinstein et al, 1977a) and β-endorphin (Rubinstein et al, 1977b) were isolated also by HPLC techniques. However, these precursor EC-proteins did not contain a Leu-enkephalin sequence.

Figure 1 shows the schema...

Table of contents

• Cover
• Half Title
• Title Page
• Copyright Page
• Preface
• Dedication
• Table of Contents
• Introduction
• Application of TSKgel SW in nucleic acid separation
• Membrane-bound hormone receptor purification and characterization by HPLC
• Determination of protein molecular weight by gel permeation chromatography equipped with low-angle laser light scattering photometer
• Gel permeation of proteins by high-performance gel chromatography in denaturing solvents
• Multiple forms of cytochrome P-450 separable by high-performance liquid chromatography
• Gel permeation studies of glycoprotein by high-performance liquid chromatography
• Gel permeation of human serum lipoprotein by HPLC
• Heterogeneity of renin: application of HPLC in studies on renin and renin binding protein
• Enzyme purification by high-performance ion-exchange liquid chromatography
• Application of HPLC for analysis and purification of catecholamine-synthesizing enzymes
• High-performance liquid chromatography for enkephalins and enkephalin-containing peptides
• Technical aspects of biochemical high-performance column liquid chromatography
• Author index
• Subject index